Substituted dihydropyrazolones and their use

ABSTRACT

The present application relates to novel substituted dihydropyrazolone derivatives, processes for their preparation, their use for treatment and/or prophylaxis of diseases and their use for the preparation of medicaments for treatment and/or prophylaxis of diseases, in particular cardiovascular and haematological diseases and kidney diseases, and for promoting wound healing.

RELATED APPLICATIONS/PATENTS AND INCORPORATION BY REFERENCE

This application claims priority to German Patent Application Number102008020113.8 filed on Apr. 23, 2008, the contents of which are herebyincorporated by reference.

The foregoing application, and all documents cited therein, and alldocuments cited or referenced therein, and all documents cited orreferenced herein, including any U.S. or foreign patents or publishedpatent applications, International patent applications, as well as, anynon-patent literature references and any manufacturer's instructions,are hereby expressly incorporated herein by reference.

BACKGROUND

1. Field of the Invention

The present application relates to novel substituted dihydropyrazolonederivatives, processes for their preparation, their use for treatmentand/or prophylaxis of diseases and their use for the preparation ofmedicaments for treatment and/or prophylaxis of diseases, in particularcardiovascular and haematological diseases and kidney diseases, and forpromoting wound healing.

2. Background of the Invention

A deficient supply of oxygen to the human organism or its componentswhich either impairs regular functioning of the organism or itscomponents due to its duration and/or its extent or causes itsfunctioning to break down completely is called hypoxia. Hypoxia can becaused by a reduction in the available oxygen in the air breathed in(for example during periods at a high altitude), by disorders inexternal respiration (for example as a result of disturbed functioningof the lungs or obstruction of the respiratory tract), by a reduction inthe cardiac output (for example in the event of cardiac insufficiency,acute right ventricular overloading with pulmonary embolism), by too lowan oxygen transport capacity of the blood (for example as a result of ananaemia or intoxication, for example with carbon monoxide), locallydemarcated by a reduced blood flow as a result of vascular occlusions(ischaemia states typically for example of the heart, the lowerextremities or the brain, diabetic macro- and microangiopathy) or alsoby an increased oxygen requirement of the tissue (for example as aresult of increased muscular activity or local inflammations) [Eder,Gedigk (ed.), Allgemeine Pathologie und pathologische Anatomie, 33rded., Springer Verlag, Berlin, 1990]

The human organism is capable to a limited extent of adapting acutelyand chronically to situations of reduced oxygen supply. In addition toan immediate response, which includes inter alia an increase in thecardiac output and respiratory output and a local dilation of bloodvessels by vegetative-nervous control mechanisms, hypoxia brings about achange in the transcription of numerous genes. The function of the geneproducts here serves to compensate the oxygen deficiency. Thus,expression of several enzymes of glycolysis and glucose transporter 1 isenhanced, as a result of which anaerobic ATP production increases andsurvival of the oxygen deficiency is rendered possible [Schmidt, Thews(ed.), Physiologie des Menschen, 27th ed., Springer Verlag, Berlin,1997; Löffler, Petrides (ed.), Biochemie und Pathobiochemie, 7th ed.,Springer Verlag, Berlin, 2003].

Hypoxia furthermore leads to enhanced expression of vascular endothelialcell growth factor, VEGF, as a result of which regeneration of bloodvessels (angiogenesis) is stimulated in hypoxic tissues. The blood flowthrough ischaemic tissue is thereby improved in the long term. Thiscounter-regulation is evidently only very inadequate in the case ofvarious cardiovascular diseases and vascular occlusion diseases[overview in: Simons and Ware, Therapeutic angiogenesis incardiovascular disease, Nat. Rev. Drug. Discov. 2 (11), 863-71 (2003)].

Furthermore, in cases of systemic hypoxia expression of the peptidehormone erythropoietin formed predominantly in the interstitialfibroblasts of the kidneys is enhanced. The formation of red blood cellsin the bone marrow is thereby stimulated, and the oxygen transportcapacity of the blood is therefore increased. This effect has been andis used by high-performance athletes in so-called high altitudetraining. A decrease in the oxygen transport capacity of the blood forexample as a result of anaemia after hemorrhaging usually causes anincrease in erythropoietin production in the kidney. With certain formsof anaemia, this regulatory mechanism may be disturbed or its normalvalue may be set lower. Thus for example in patients suffering fromrenal insufficiency, erythropoietin is indeed produced in the kidneyparenchyma, but in significantly reduced amounts with respect to theoxygen transport capacity of the blood, which results in so-called renalanaemia. Renal anaemia in particular, but also anemias caused by tumoursand HIV infection are conventionally treated by parenteraladministration of recombinant human erythropoietin (rhEPO). Noalternative therapy with an orally available medicament currently existsfor this expensive therapy [overview in: Eckardt, The potential oferythropoietin and related strategies to stimulate erythropoiesis, Curr.Opin. Investig. Drugs 2(8), 1081-5 (2001); Berns, Should the targethemoglobin for patients with chronic kidney disease treated witherythropoietic replacement therapy be changed?, Semin. Dial. 18 (1),22-9 (2005)]. Recent studies demonstrate that, in addition to itserythropoiesis-increasing action, erythropoietin also has a protective(anti-apoptotic) action, which is independent thereof, on hypoxictissue, in particular the heart and the brain.

Furthermore, according to recent studies therapy with erythropoietinreduces the average severity of morbidity in patients with cardiacinsufficiency [overviews in: Caiola and Cheng, Use of erythropoietin inheart failure management, Ann. Pharmacother. 38 (12), 2145-9 (2004);Katz, Mechanisms and treatment of anemia in chronic heart failure,Congest. Heart. Fail. 10 (5), 243-7 (2004)].

The genes described above which are induced by hypoxia have the commonfeature that the increase in their expression under hypoxia is caused bythe so-called hypoxia-inducible transcription factor (HIF). HIF is aheterodimeric transcription factor which comprises an alpha and a betasubunit. Three HIF alpha isoforms have been described, of which HIF-1alpha and HIF-2 alpha are highly homologous and are of importance forhypoxia-induced gene expression. While the beta subunit (of which 2isoforms have been described), which is also called ARNT (arylhydrocarbon receptor nuclear translocator), is expressed constitutively,expression of the alpha subunit depends on the oxygen content in thecell. Under normoxia, the HIF alpha protein is poly-ubiquitinized andthen degraded proteasomally. Under hypoxia this degradation isinhibited, so that HIF alpha dimerizes with ARNT and can activate itstarget genes. The HIF dimer bonds here to so-called hypoxia-responsibleelements (HRE) in the regulatory sequences of its target genes. The HREare defined by a consensus sequence. Functional HRE have been detectedin the regulatory elements of numerous hypoxia-induced genes [overviewsin: Semenza, Hypoxia-inducible factor 1: oxygen homeostasis and diseasepathophysiology, Trends Mol. Med. 7 (8), 345-50 (2001); Wenger andGassmann, Oxygen(es) and the hypoxia-inducible factor-1, Biol. Chem. 378(7), 609-16 (1997)].

The molecular mechanism on which this regulation of HIF alpha is basedhas been clarified by the works of several independent groups ofresearchers. The mechanism is conserved from species to species: HIFalpha is hydroxylated by a subclass of oxygen-dependent prolyl4-hydroxylases, called PHD or EGLN, on two specific prolyl radicals(P402 and P564 of the human HIF-1 alpha subunit). The HIF prolyl4-hydroxylases are iron-dependent, 2-oxoglutarate-convertingdioxygenases [Epstein et al., C. elegans EGL-9 and mammalian homologsdefine a family of dioxygenases that regulate HIF by prolylhydroxylation, Cell 107 (1), 43-54 (2001); Bruick and McKnight, Aconserved family of prolyl-4-hydroxylases that modify HIF, Science 294(5545), 1337-40 (2001); Ivan et al., Biochemical purification andpharmacological inhibition of a mammalian prolyl hydroxylase acting onhypoxia-inducible factor, Proc. Natl. Acad. Sci. U.S.A. 99 (21),13459-64 (2002)]. The enzymes were annotated as prolyl hydroxylases forthe first time in 2001 [Aravind and Koonin, The DNA-repair protein AlkB,EGL-9, and leprecan define new families of 2-oxoglutarate- andiron-dependent dioxygenases, Genome Biol. 2 (3), research0007.1-0007.8,Epub 2001 Feb. 19].

The pVHL tumour suppressor protein, which together with elongin B and Cforms the so-called VBC complex, which adapts the HIF alpha subunit toan E3 ubiquitin ligase, bonds to the prolyl-hydroxylated HIF alphasubunit. Since the prolyl 4-hydroxylation of the HIF alpha subunit andits subsequent degradation takes place as a function of theintracellular concentration of oxygen, HIF prolyl 4-hydroxylases havealso been called a cellular oxygen sensor. Three isoforms of theseenzymes have been identified: EGLN1/PHD2, EGLN2/PHD1 and EGLN3/PHD3. Twoof these enzymes (EGLN2/PHD1 and EGLN3/PHD3) are inducedtranscriptionally even under hypoxia and are possibly responsible forthe lowering of the HIF alpha levels to be observed under chronichypoxia [overview in: Schofield and Ratcliffe, Oxygen sensing by HIFhydroxylases, Nat. Rev. Mol. Cell. Biol. 5 (5), 343-54 (2004)].

Selective pharmacological inhibition of HIF prolyl 4-hydroxylases bringsabout the increase in the gene expression of HIF-dependent target genesand is therefore beneficial for the therapy of numerous diseasesyndromes. In the case of diseases of the cardiovascular system inparticular, an improvement in the course of the diseases is to beexpected from induction of new blood vessels and the change in themetabolic situation of ischaemic organs from aerobic to anaerobic ATPproduction. An improvement in the vascularization of chronic woundspromotes the healing process, especially in the case of poorly healingulcera cruris and other chronic skin wounds. The induction of endogenouserythropoietin in certain disease forms, in particular in patients withrenal anaemia, is likewise a therapeutic goal to be aimed for.

The HIF prolyl 4-hydroxylase inhibitors described hitherto in thescientific literature do not meet the requirements to be imposed on amedicament. These are either competitive oxoglutarate analogues (such asfor example N-oxalylglycine), which are characterized by their very lowaction potency, and therefore in in vivo models have as yet shown noaction in the sense of an induction of HIF target genes. Or they areiron-complexing agents (chelators), such as desferroxamine, which act asnon-specific inhibitors of iron-containing dioxygenases and, althoughthey bring about an induction of the target genes, such as for exampleerythropoietin, in vivo, evidently counteract erythropoiesis bycomplexing of the available iron.

2-Heteroaryl-4-aryl-1,2-dihydropyrazolones having a bactericidal and/orfungicidal action are disclosed in EP 165 448 and EP 212 281. The use of2-heteroaryl-4-aryl-1,2-dihydropyrazolones as lipoxygenase inhibitorsfor treatment of respiratory tract, cardiovascular and inflammatorydiseases is claimed in EP 183 159. 2,4-Diphenyl-1,2-dihydropyrazoloneshaving a herbicidal activity are described in DE 2 651 008. Thepreparation and pharmacological properties of certain2-pyridyl-1,2-dihydropyrazolones are reported in Helv. Chim. Acta 49(1), 272-280 (1966). WO 96/12706, WO 00/51989 and WO 03/074550 claimcompounds having a dihydropyrazolone partial structure for treatment ofvarious diseases, and hydroxy- or alkoxy-substituted bipyrazoles fortreatment of neuropsychiatric diseases are disclosed in WO 2006/101903.Heteroaryl-substituted pyrazole derivatives for treatment of pain andvarious CNS diseases are furthermore described in WO 03/051833 and WO2004/089303. WO 2006/114213 has meanwhile disclosed2,4-dipyridyl-1,2-dihydropyrazolones as inhibitors of HIF prolyl4-hydroxylases.

The X-ray crystal structure of the compound3-methyl-1-(pyridin-2-yl)-4-(1-pyridin-2-yl-3-methyl-1H-pyrazol-5-yl)-2H-3-pyrazolin-5(1H)-one(other name:5,5′-dimethyl-2,2′-dipyridin-2-yl-1′,2′-dihydro-2H,3′H-3,4′-bipyrazol-3′-one)is reported in Acta Crystallogr., Section E: Structure Reports OnlineE57 (11), o1126-o1127 (2001) [Chem. Abstr. 2001:796190]. The synthesisof certain3′,5-dimethyl-2-phenyl-1′-(1,3-thiazol-2-yl)-1′H,2H-3,4′-bipyrazol-5′-olderivatives is described in Indian J. Heterocyclic Chem. 3 (1), 5-8(1993) [Chem. Abstr. 1994:323362]. The preparation and tautomerism ofindividual 4-(pyrazol-5-yl)pyrazolin-5-one derivatives is reported in J.Heterocyclic Chem. 27 (4), 865-870 (1990) [Chem. Abstr. 1991:428557]. Atherapeutic use has not hitherto been described for the compoundsmentioned in these publications. The compound2-tert-butyl-1′-[4-(4-chlorophenyl)-1,3-thiazol-2-yl]-3′,5-dimethyl-1′H,2H-3,4′-bipyrazol-5′-olis listed as a test example in WO 2007/008541.

SUMMARY

An object of the present invention is to provide novel compounds whichcan be employed for treatment of diseases, in particular cardiovascularand haematological diseases.

In the context of the present invention, compounds are now describedwhich act as specific inhibitors of HIF prolyl 4-hydroxylases and on thebasis of this specific action mechanism bring about in vivo, afterparenteral or oral administration, the induction of HIF target genes,such as e.g. erythropoietin, and the biological processes therebycaused, such as e.g. erythropoiesis.

The present invention provides compounds of the formula

-   in which-   X represents N or CH,-   R¹ represents hydrogen or cyano,-   R² represents a saturated 4- to 7-membered heterocyclyl radical    which is attached via a nitrogen atom,    -   where the heterocyclyl radical may be substituted by a        substituent, where the substituent is selected from the group        consisting of hydroxyl, hydroxycarbonyl, C₁-C₃-alkyl,        C₁-C₃-alkylamino and C₃-C₆-cycloalkyl,    -   or    -   where the heterocyclyl radical may be substituted by 1 to 4        fluorine substituents,-   and salts, solvates and solvates of the salts thereof.

DETAILED DESCRIPTION

Compounds according to the invention are the compounds of the formula(I) and their salts, solvates and solvates of the salts, and thecompounds included in the formula (I) and mentioned in the following asembodiment examples and their salts, solvates and solvates of the salts,where the compounds included in the formula (I) and mentioned in thefollowing are not already salts, solvates and solvates of the salts.

The compounds according to the invention can exist in stereoisomericforms (enantiomers, diastereomers), depending on their structure. Theinvention therefore includes the enantiomers or diastereomers and theirparticular mixtures. The stereoisomerically uniform constituents can beisolated from such mixtures of enantiomers and/or diastereomers in aknown manner.

Where the compounds according to the invention can occur in tautomericforms, the present invention includes all the tautomeric forms.

Preferred salts in the context of the present invention arephysiologically acceptable salts of the compounds according to theinvention. Salts which are not themselves suitable for pharmaceuticaluses but can be used, for example, for isolation or purification of thecompounds according to the invention are also included.

Physiologically acceptable salts of the compounds according to theinvention include acid addition salts of mineral acids, carboxylic acidsand sulphonic acids, for example salts of hydrochloric acid, hydrobromicacid, sulphuric acid, phosphoric acid, methanesulphonic acid,ethanesulphonic acid, toluenesulphonic acid, benzenesulphonic acid,naphthalenedisulphonic acid, acetic acid, trifluoroacetic acid,propionic acid, lactic acid, tartaric acid, malic acid, citric acid,fumaric acid, maleic acid and benzoic acid.

Physiologically acceptable salts of the compounds according to theinvention also include salts of conventional bases, such as, by way ofexample and preferably, alkali metal salts (for example sodium andpotassium salts), alkaline earth metal salts (for example calcium andmagnesium salts) and ammonium salts derived from ammonia or organicamines having 1 to 16 C atoms, such as, by way of example andpreferably, ethylamine, diethylamine, triethylamine,ethyldiisopropylamine, monoethanolamine, diethanolamine,triethanolamine, dicyclohexylamine, dimethylaminoethanol, procaine,dibenzylamine, N-methylmorpholine, arginine, lysine, ethylenediamine andN-methylpiperidine.

Solvates in the context of the invention are described as those forms ofthe compounds according to the invention which form a complex in thesolid or liquid state by coordination with solvent molecules. Hydratesare a specific form of solvates, in which the coordination takes placewith water. Hydrates are preferred solvates in the context of thepresent invention.

The present invention moreover also includes prodrugs of the compoundsaccording to the invention. The term “prodrugs” includes compounds whichthemselves can be biologically active or inactive, but are converted(for example metabolically or hydrolytically) into compounds accordingto the invention during their dwell time in the body.

In the context of the present invention, the substituents have thefollowing meaning, unless specified otherwise:

Alkyl per se and “alkyl” in alkylamino represent a straight-chain orbranched alkyl radical having 1 to 3 carbon atoms, by way of example andby way of preference methyl, ethyl, n-propyl, isopropyl.

Alkylamino represents an alkylamino radical having one or two (selectedindependently of one another) alkyl substituents, by way of example andby way of preference methylamino, ethylamino, n-propylamino,isopropylamino, N,N-dimethylamino, N,N-diethylamino,N-ethyl-N-methylamino, N-methyl-N-n-propylamino andN-isopropyl-N-n-propylamino. C₁-C₃-Alkylamino, for example, represents amonoalkylamino radical having 1 to 3 carbon atoms or represents adialkylamino radical having 1 to 3 carbon atoms each per alkylsubstituent.

Cycloalkyl represents a monocyclic cycloalkyl group having generally 3to 6 carbon atoms; cycloalkyl radicals which may be mentioned by way ofexample and by way of preference are cyclopropyl, cyclobutyl,cyclopentyl and cyclohexyl.

A saturated 4- to 7-membered heterocyclyl radical which is attached viaa nitrogen atom represents a monocyclic saturated heterocyclic radicalhaving 4 to 7 ring atoms which contain one nitrogen atom via which it isattached and up to 2, preferably up to one, further heteroatom(s) and/ora heterogroup selected from the group consisting of N, O, S, SO, SO₂,where a nitrogen atom may also form an N-oxide. Preference is given to4- to 7-membered monocyclic saturated heterocyclyl radicals having up toone further heteroatom from the group consisting of O, N and S, by wayof example and by way of preference azetidin-1-yl, pyrrolin-1-yl,piperidin-1-yl, morpholin-4-yl, thiomorpholin-4-yl, piperazin-1-yl,1,2-oxazinan-2-yl, 1,4-oxazepan-4-yl, 1,4-thiazepan-4-yl.

Preference is given to compounds of the formula (I), in which

-   X represents N or CH,-   R¹ represents hydrogen or cyano,-   R² represents a saturated 4- to 7-membered heterocyclyl radical    which is attached via a nitrogen atom,    -   where the heterocyclyl radical is substituted by 1 to 4 fluorine        substituents,-   or-   R² represents piperazin-1-yl,    -   where piperazin-1-yl is substituted by one substituent, where        the substituent is selected from the group consisting of        C₃-C₆-cycloalkyl,-   or-   R² represents azetidin-1-yl,    -   where azetidin-1-yl is substituted by one substituent, where the        substituent is selected from the group consisting of        hydroxycarbonyl, C₁-C₃-alkyl, C₁-C₃-alkylamino and        C₃-C₆-cycloalkyl,-   or-   R² represents 1,2-oxazinan-2-yl or 1,4-oxazepan-4-yl,-   and salts, solvates and solvates of the salts thereof.

Preference is given to compounds of the formula (I) in which

-   X represents N or CH,-   R¹ represents hydrogen or cyano,-   R² represents azetidin-1-yl, pyrrolin-1-yl or piperidin-1-yl,    -   where azetidin-1-yl, pyrrolin-1-yl and piperidin-1-yl are        substituted by 1 to 4 fluorine substituents,-   or-   R² represents piperazin-1-yl,    -   where piperazin-1-yl is substituted in the 4-position by one        substituent, where the substituent is selected from the group        consisting of C₃-C₆-cycloalkyl,-   or-   R² represents azetidin-1-yl,    -   where azetidin-1-yl is substituted in the 3-position by one        substituent, where the substituent is selected from the group        consisting of hydroxycarbonyl, methyl and dimethylamino,-   or-   R² represents 1,2-oxazinan-2-yl or 1,4-oxazepan-4-yl,-   and salts, solvates and solvates of the salts thereof.

Preference is also given to compounds of the formula (I) in which

-   X represents N or CH,-   R¹ represents hydrogen or cyano,-   R² represents a saturated 4- to 7-membered heterocyclyl radical    which is attached via a nitrogen atom,    -   where the heterocyclyl radical is substituted by 1 to 4 fluorine        substituents,-   and salts, solvates and solvates of the salts thereof.

Preference is also given to compounds of the formula (I) in which

-   X represents N or CH,-   R¹ represents hydrogen or cyano,-   R² represents azetidin-1-yl, pyrrolin-1-yl or piperidin-1-yl,    -   where azetidin-1-yl, pyrrolin-1-yl and piperidin-1-yl are        substituted by 1 to 4 fluorine substituents,-   and salts, solvates and solvates of the salts thereof.

Preference is also given to compounds of the formula (I) in which

-   X represents N or CH,-   R¹ represents hydrogen or cyano,-   R² represents azetidin-1-yl, pyrrolin-1-yl or piperidin-1-yl,    -   where azetidin-1-yl, pyrrolin-1-yl and piperidin-1-yl are        substituted by 2 fluorine substituents, where these substituents        are attached to the same carbon atom,-   and salts, solvates and solvates of the salts thereof.

Preference is also given to compounds of the formula (I) in which

-   X represents N or CH,-   R¹ represents hydrogen or cyano,-   R² represents piperazin-1-yl,    -   where piperazin-1-yl is substituted by one substituent, where        the substituent is selected from the group consisting of        C₃-C₆-cycloalkyl,-   and salts, solvates and solvates of the salts thereof.

Preference is also given to compounds of the formula (I) in which

-   X represents N or CH,-   R¹ represents hydrogen or cyano,-   R² represents piperazin-1-yl,    -   where piperazin-1-yl is substituted in the 4-position by one        substituent, where the substituent is selected from the group        consisting of C₃-C₆-cycloalkyl,-   and salts, solvates and solvates of the salts thereof.

Preference is also given to compounds of the formula (I) in which

-   X represents N or CH,-   R¹ represents hydrogen or cyano,-   R² represents azetidin-1-yl,    -   where azetidin-1-yl is substituted by one substituent, where the        substituent is selected from the group consisting of        hydroxycarbonyl, C₁-C₃-alkyl, C₁-C₃-alkylamino and        C₃-C₆-cycloalkyl,-   and salts, solvates and solvates of the salts thereof.

Preference is also given to compounds of the formula (I) in which

-   X represents N or CH,-   R¹ represents hydrogen or cyano,-   R² represents azetidin-1-yl,    -   where azetidin-1-yl is substituted by one substituent, where the        substituent is selected from the group consisting of        hydroxycarbonyl, methyl and dimethylamino,-   and salts, solvates and solvates of the salts thereof.

Preference is also given to compounds of the formula (I) in which

-   X represents N or CH,-   R¹ represents hydrogen or cyano,-   R² represents azetidin-1-yl,    -   where azetidin-1-yl is substituted in the 3-position by one        substituent, where the substituent is selected from the group        consisting of hydroxycarbonyl, methyl and dimethylamino,-   and salts, solvates and solvates of the salts thereof.

Preference is also given to compounds of the formula (I) in which

-   X represents N or CH,-   R¹ represents hydrogen or cyano,-   R² represents 1,2-oxazinan-2-yl or 1,4-oxazepan-4-yl.

Preference is also given to compounds of the formula (I) in which Xrepresents N.

Preference is also given to compounds of the formula (I) in which R¹represents hydrogen.

Preference is also given to compounds of the formula (I) in which R¹represents cyano.

Preference is also given to compounds of the formula (I) in which R²represents 4-cyclobutyl-piperazin-1-yl.

Preference is also given to compounds of the formula (I) in which

-   X represents N or CH,-   R¹ represents cyano,-   R² represents a saturated 4- to 7-membered heterocyclyl radical    which is attached via a nitrogen atom,    -   where the heterocyclyl radical is substituted by one        substituent, where the substituent is selected from the group        consisting of hydroxy, hydroxycarbonyl, C₁-C₃-alkyl,        C₁-C₃-alkylamino and C₃-C₆-cycloalkyl,    -   or    -   where the heterocyclyl radical is substituted by 1 to 4 fluorine        substituents,-   and salts, solvates and solvates of the salts thereof.

The radical definitions given in detail in the particular combinationsor preferred combinations of radicals are also replaced as desired byradical definitions of other combinations, independently of theparticular radical combinations given.

Combinations of two or more of the abovementioned preferred ranges arevery particularly preferred.

The 1,2-dihydropyrazol-3-one derivatives of the formula (I) according tothe invention can also be in the tautomeric 1H-pyrazol-5-ol form (I′)(see following Scheme 1); the two tautomeric forms are expresslyincluded in the present invention.

The invention also provides a process for the preparation of thecompounds of the formula (I), or salts, solvates and solvates of thesalts thereof, wherein, according to process

[A] compounds of the formula

in which R¹ has the meaning given above, andZ¹ represents methyl or ethyl,are reacted in an inert solvent, if appropriate in the presence of anacid, with a compound of the formula

in which R has the meaning given above,to give compounds of the formula

in which Z¹, R¹ and R² have the meaning given above,which, already under these reaction conditions or in a subsequentreaction step under the influence of a base, cyclize to give thecompounds of the formula (I),and the compounds of the formula (I) are, if appropriate with theappropriate (i) solvents and/or (ii) bases or acids, converted intotheir salts, their solvates, or the solvates of their salts,or[B] compounds of the formula

in which Z¹ and R¹ have the meaning given above,are condensed with a compound of the formula

in whichZ² represents methyl or ethyl,to give compounds of the formula

in which Z¹ and R¹ have the meaning given above,and then reacted in the presence of an acid with a compound of theformula (III) to give compounds of the formula (IV), which, alreadyunder these reaction conditions or in a subsequent reaction step underthe influence of a base, cyclize to give the compounds of the formula(I),and the compounds of the formula (I) are, if appropriate with theappropriate (i) solvents and/or (ii) bases or acids, converted intotheir salts, their solvates, or the solvates of their salts,or[C] the compound of the formula

is, in water as the solvent in a one-pot process, reacted initially withcompounds of the formulaR²—H  (IX),in which R² has the meaning given above,and then with compounds of the formula (VII) to give compounds of theformula (I),and the compounds of the formula (I) are, if appropriate with theappropriate (i) solvents and/or (ii) bases or acids, converted intotheir salts, their solvates, or the solvates of their salts.

The free base of the salts can be obtained by reacting the salts of thecompounds or solvates of the salts of the compounds with a base.

Suitable bases are alkali metal hydroxides such as, for example, sodiumhydroxide or potassium hydroxide, alkali metal or alkaline earth metalcarbonates, such as sodium carbonate, potassium carbonate, calciumcarbonate or caesium carbonate, or aqueous ammonia solution.

In an alternative process, the free base of the salts can be obtained,for example, by chromatography on a reversed-phase column using anacetonitrile/water gradient with addition of a base, in particular byusing an RP18 Phenomenex Luna C18(2) column and the base diethylamine.

The invention furthermore provides a process for preparing the compoundsof the formula (I) or solvates thereof where salts of the compounds orsolvates of the salts of the compounds are converted by reaction with abase or by chromatography with addition of a base into the compounds.

Further compounds according to the invention can optionally also beprepared by conversions of functional groups of individual substituents,in particular those listed under R¹ and R², starting from the compoundsof the formula (I) obtained by the above processes. These conversionsare carried out by conventional methods known to the person skilled inthe art and include, for example, reactions such as nucleophilic orelectrophilic substitution, oxidation, reduction, hydrogenation,transition metal-catalysed coupling reactions, alkylation, acylation,amination, esterification, ester cleavage, etherification, ethercleavage, formation of carboxamides, sulphonamides, carbamates andureas, and the introduction and removal of temporary protective groups.

Suitable inert solvents for the process steps (II)+(III)→(IV),(VII)+(III)→(IV) and (IV)→(I) are in particular ethers, such as diethylether, methyl tert-butyl ether, 1,2-dimethoxyethane, tetra-hydrofuranand dioxane, or alcohols, such as methanol, ethanol, n-propanol,isopropanol, n-butanol and tert-butanol, or water or mixtures of thesolvents or a mixture of a solvent with water. Preference is given tousing methanol, ethanol, tetrahydrofuran or water.

The process step (V)+(VI)→(VII) is preferably carried out in the solventdimethylformamide or else in the presence of an excess of (VI) withoutfurther solvent. If appropriate, the reaction may also be carried out inan advantageous manner under microwave irradiation. The reaction isgenerally carried out in a temperature range of from +20° C. to +150°C., preferably at from +80° C. to +120° C. [cf. also J. P. Bazureau etal., Synthesis 1998, 967; ibid. 2001 (4), 581].

If appropriate, the process steps (II)+(III)→(IV) and (VII)+(III)→(IV)can be carried out in an advantageous manner with addition of an acid.Suitable for this purpose are customary inorganic or organic acids, suchas, for example, hydrogen chloride, acetic acid, trifluoroacetic acid,methane-sulphonic acid, p-toluenesulphonic acid or camphor-10-sulphonicacid. Preference is given to using acetic acid or in particulartrifluoroacetic acid or p-toluenesulphonic acid.

The reaction (II)+(III)→(IV) is generally carried out in a temperaturerange of from 0° C. to +100° C., preferably from +10° C. to +50° C. Thereaction (VII)+(III)→(IV) is generally carried out in a temperaturerange of from +20° C. to +120° C., preferably from +50° C. to +100° C.

The process sequences (II)+(III)→(IV)→(I) and (VII)+(III)→(IV)→(I) canbe carried out as a two-step reaction or else as a one-pot reaction,without isolation of the intermediate (IV). Especially suitable for thelatter variant is the reaction of the components under microwaveirradiation; here, the reaction is generally carried out in atemperature range of from +50° C. to +200° C., preferably from +100° C.to +180° C.

There may be partial ring closure to (I) even during the preparation of(IV); in this case, the cyclization can, if required, be brought tocompletion by treating the reaction mixture in situ with a base.

Suitable bases for such a separate cyclization step (IV)→(I) arecustomary inorganic or organic bases. These include in particular alkalimetal hydroxides, such as, for example, sodium hydroxide or potassiumhydroxide, alkali metal carbonates or alkaline earth metal carbonates,such as sodium carbonate, potassium carbonate, calcium carbonate orcaesium carbonate, alkali metal alkoxides, such as sodium methoxide orpotassium methoxide, sodium ethoxide or potassium ethoxide or sodiumtert-butoxide or potassium tert-butoxide, or alkali metal hydrides, suchas sodium hydride. Preference is given to using sodium methoxide orsodium ethoxide.

The base-induced reaction (IV)→(I) is generally carried out in atemperature range of from 0° C. to +60° C., preferably at from 0° C. to+30° C.

The reaction (VIII)+(IX)+(VII)→(I) is generally carried out using from1.1 to 2.0 equivalents of (IX) per equivalent of (VIII), if appropriatein the presence of from 1.1 to 2.0 equivalents of a base. Preferred isthe reaction with from 1.1 to 1.5 equivalents of (IX).

Suitable bases for the reaction (VIII)+(IX)+(VII)→(I) are customaryinorganic or organic bases. These include in particular alkali metalhydroxides, such as, for example, sodium hydroxide or potassiumhydroxide, or amine bases, such as, for example,N-ethyl-N-(propan-2-yl)propane-2-amine. Preference is given toN-ethyl-N-(propan-2-yl)propane-2-amine.

The reaction (VIII)+(IX)+(VII)→(I) is generally carried out in atemperature range of from +20° C. to +100° C., preferably at from +70°C. to +100° C.

All process steps can be carried out at atmospheric, elevated or reducedpressure (for example from 0.5 to 5 bar). In general, the reactions arecarried out at atmospheric pressure.

The compounds of the formula (II) can be prepared from compounds of theformula (V) by methods customarily used in the literature for theC-acylation of carboxylic esters. The compounds of the formulae (III),(V), (VI), (VIII) and (IX) are commercially available, known from theliterature or can be prepared analogously to processes described in theliterature.

The preparation of the compounds according to the invention can beillustrated by Reaction Scheme 2 below:

[a): DMF, 16 h, +100° C.; b): ethanol, trifluoroacetic acid, +78° C.;c): NaOEt, ethanol, 1 h, RT].

The compounds according to the invention show an unforeseeable, valuablepharmacological action spectrum.

They are therefore suitable for use as medicaments for treatment and/orprophylaxis of diseases in humans and animals.

The compounds according to the invention are distinguished as specificinhibitors of HIF prolyl 4-hydroxylases.

On the basis of their pharmacological properties, the compoundsaccording to the invention can be employed for treatment and/orprophylaxis of cardiovascular diseases, in particular cardiacinsufficiency, coronary heart disease, angina pectoris, myocardialinfarction, stroke, arteriosclerosis, essential, pulmonary and malignanthypertension and peripheral arterial occlusive disease.

The compounds according to the invention are furthermore suitable fortreatment and/or prophylaxis of blood formation disorders, such as e.g.idiopathic anaemias, renal anaemia and anaemias accompanying a tumourdisease (in particular an anaemia induced by chemotherapy), an infection(in particular HIV infection) or another inflammatory disease, such ase.g. rheumatoid arthritis. The compounds according to the invention aremoreover suitable for supporting treatment of anaemias as a result ofblood loss, iron deficiency anaemia, vitamin deficiency anaemia (e.g. asa result of vitamin B12 deficiency or as a result of folic aciddeficiency), hypoplastic and aplastic anaemia or haemolytic anaemia, orfor supporting treatment of anaemias as a result of iron utilizationdisorders (sideroachrestic anaemia) or anaemias as a result of otherendocrine disorders (e.g. hypothyroidosis).

The compounds are furthermore suitable for increasing the haematocritwith the aim of obtaining blood for autodonation of blood beforeoperations.

The compounds according to the invention can moreover be used fortreatment and/or prophylaxis of operation-related states of ischaemiaand consecutive symptoms thereof after surgical interventions, inparticular interventions on the heart using a heart-lung machine (e.g.bypass operations, heart valve implants), interventions on the carotidarteries, interventions on the aorta and interventions with instrumentalopening or penetration of the skull cap. The compounds are furthermoresuitable for general treatment and/or prophylaxis in the event ofsurgical interventions with the aim of accelerating wound healing andshortening the convalescence time.

The compounds are moreover suitable for treatment and prophylaxis ofconsecutive symptoms of acute and protracted ischaemic states of thebrain (e.g. stroke, birth asphyxia).

The compounds can furthermore be employed for treatment and/orprophylaxis of cancer and for treatment and/or prophylaxis of animpairment in the state of health occurring in the course of treatmentof cancer, in particular after therapy with cytostatics, antibiotics andirradiations.

The compounds are furthermore suitable for treatment and/or prophylaxisof diseases of the rheumatic type and other disease forms to be countedas autoimmune diseases, and in particular for treatment and/orprophylaxis of an impairment in the state of health occurring in thecourse of medicamentous treatment of such diseases.

The compounds according to the invention can moreover be employed fortreatment and/or prophylaxis of diseases of the eye (e.g. glaucoma), thebrain (e.g. Parkinson's disease, Alzheimer's disease, dementia, chronicpain sensation), of chronic kidney diseases, renal insufficiency andacute renal failure and for promoting wound healing.

The compounds are moreover suitable for treatment and/or prophylaxis ofgeneral physical weakness, up to cachexia, in particular occurring to anincreased extent at a more elderly age.

The compounds are furthermore suitable for treatment and/or prophylaxisof sexual dysfunction.

The compounds are moreover suitable for treatment and/or prophylaxis ofdiabetes mellitus and its consecutive symptoms, such as e.g. diabeticmacro- and microangiopathy, diabetic nephropathy and neuropathy.

The compounds according to the invention are moreover suitable fortreatment and/or prophylaxis of fibrotic diseases e.g. of the heart, thelungs and the liver.

In particular, the compounds according to the invention are alsosuitable for prophylaxis and treatment of retinopathy in prematurebabies (retinopathia prematurorum).

The present invention moreover provides the use of the compoundsaccording to the invention for treatment and/or prevention of diseases,in particular the abovementioned diseases.

The present invention moreover provides the use of the compoundsaccording to the invention for the preparation of a medicament fortreatment and/or prevention of diseases, in particular theabovementioned diseases.

The present invention moreover provides a method for treatment and/orprevention of diseases, in particular the abovementioned diseases, usingan active amount of at least one of the compounds according to theinvention.

The compounds according to the invention can be employed by themselvesor, if required, in combination with other active compounds. The presentinvention moreover provides medicaments comprising at least one of thecompounds according to the invention and one or more further activecompounds, in particular for treatment and/or prevention of theabovementioned diseases. Suitable active compounds in the combinationwhich may be mentioned by way of example and preferably are: ACEinhibitors, angiotensin II receptor antagonists, beta receptor blockers,calcium antagonists, PDE inhibitors, mineralocorticoid receptorantagonists, diuretics, aspirin, iron supplements, vitamin B12 and folicacid supplements, statins, digitalis (digoxin) derivatives, tumourchemotherapeutics and antibiotics.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an ACE inhibitor,such as, by way of example and preferably, enalapril, captopril,lisinopril, ramipril, delapril, fosinopril, quinopril, perindopril ortrandopril.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an angiotensin AIIantagonist, such as, by way of example and preferably, losartan,candesartan, valsartan, telmisartan or embusartan.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a beta receptorblocker, such as, by way of example and preferably, propranolol,atenolol, timolol, pindolol, alprenolol, oxprenolol, penbutolol,bupranolol, metipranolol, nadolol, mepindolol, carazalol, sotalol,metoprolol, betaxolol, celiprolol, bisoprolol, carteolol, esmolol,labetalol, carvedilol, adaprolol, landiolol, nebivolol, epanolol orbucindolol.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a calcium antagonist,such as, by way of example and preferably, nifedipine, amlopidine,verapamil or diltiazem.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a phosphodiesterase(PDE) inhibitor, such as, by way of example and preferably, milrinone,aminone, pimobendan, cilostazol, sildenafil, vardenafil or tadalafil.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a mineralocorticoidreceptor antagonist, such as, by way of example and preferably,spironolactone, eplerenone, canrenone or potassium canrenoate.

In a preferred embodiment of the invention the compounds according tothe invention are administered in combination with a diuretic, such as,by way of example and preferably, furosemide, bumetamide, torsemide,bendroflumethiazide, chlorothiazide, hydrochlorothiazide,hydroflumethiazide, methylclothiazide, polythiazide,trichloromethiazide, chlorthalidone, indapamide, metolazone,quinethazone, acetazolamide, dichlorphenamide, methazolamide, glycerine,isosorbide, mannitol, amiloride or triamteren.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an HMG-CoA reductaseinhibitor from the class of statins, such as, by way of example andpreferably, lovastatin, simvastatin, pravastatin, fluvastatin,atorvastatin, rosuvastatin, cerivastatin or pitavastatin.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a tumourchemotherapeutic, by way of example and preferably from the groupconsisting of platinum complexes, such as e.g. cisplatin andcarboplatin, the alkylating agents, such as e.g. cyclophosphamide andchlorambucil, the antimetabolites, such as e.g. 5-fluorouracil andmethotrexate, the topoisomerase inhibitors, such as e.g. etoposide andcamptothecin, the antibiotics, such as e.g. doxorubicin anddaunorubicin, or the kinase inhibitors, such as e.g. sorafenib andsunitinib.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an antibiotic, by wayof example and preferably from the group consisting of penicillins,cephalosporins or quinolones, such as e.g. ciprofloxacin andmoxifloxacin.

The present invention moreover provides medicaments which comprise atleast one compound according to the invention, conventionally togetherwith one or more inert, non-toxic, pharmaceutically suitable auxiliarysubstances, and the use thereof for the above-mentioned purposes.

The compounds according to the invention can act systemically and/orlocally. They can be administered in a suitable manner for this purpose,such as e.g. orally, parenterally, pulmonally, nasally, sublingually,lingually, buccally, rectally, dermally, transdermally, conjunctivally,optically or as an implant or stent.

The compounds according to the invention can be administered in suitableadministration forms for these administration routes.

Administration forms which function according to the prior art, releasethe compounds according to the invention rapidly and/or in a modifiedmanner and comprise the compounds according to the invention incrystalline and/or amorphized and/or dissolved form are suitable fororal administration, such as e.g. tablets (non-coated or coated tablets,for example coatings which are resistant to gastric juice or dissolve ina delayed manner or are insoluble and control the release of thecompound according to the invention), tablets or films/oblates,films/lyophilizates or capsules which disintegrate rapidly in the oralcavity (for example hard or soft gelatin capsules), sugar-coatedtablets, granules, pellets, powders, emulsions, suspensions, aerosols orsolutions.

Parenteral administration can be effected with bypassing of anabsorption step (e.g. intravenously, intraarterially, intracardially,intraspinally or intralumbarly) or with inclusion of an absorption (e.g.intramuscularly, subcutaneously, intracutaneously, percutaneously orintraperitoneally). Administration forms which are suitable forparenteral administration are, inter alia, injection and infusionformulations in the form of solutions, suspensions, emulsions,lyophilizates or sterile powders.

For the other administration routes e.g. inhalation medicament forms(inter alia powder inhalers, nebulizers), nasal drops, solutions orsprays, tablets, films/oblates or capsules for lingual, sublingual orbuccal administration, suppositories, ear or eye preparations, vaginalcapsules, aqueous suspensions (lotions, shaking mixtures), lipophilicsuspensions, ointments, creams, transdermal therapeutic systems (e.g.patches), milk, pastes, foams, sprinkling powders, implants or stentsare suitable.

Oral and parenteral administration are preferred, in particular oral andintravenous administration.

The compounds according to the invention can be converted into theadministration forms mentioned. This can be effected in a manner knownper se by mixing with inert, non-toxic, pharmaceutically suitableauxiliary substances. These auxiliary substances include inter aliacarrier substances (for example microcrystalline cellulose, lactose,mannitol), solvents (e.g. liquid polyethylene glycols), emulsifiers anddispersing or wetting agents (for example sodium dodecyl sulphate,polyoxysorbitan oleate), binders (for example polyvinylpyrrolidone),synthetic and natural polymers (for example albumin), stabilizers (e.g.antioxidants, such as, for example, ascorbic acid), dyestuffs (e.g.inorganic pigments, such as, for example, iron oxides) and flavourand/or smell correctants.

In general, it has proved advantageous in the case of parenteraladministration to administer amounts of from about 0.001 to 1 mg/kg,preferably about 0.01 to 0.5 mg/kg of body weight to achieve effectiveresults. In the case of oral administration the dosage is about 0.01 to100 mg/kg, preferably about 0.01 to 20 mg/kg and very particularlypreferably 0.1 to 10 mg/kg of body weight.

Nevertheless it may be necessary to deviate from the amounts mentioned,and in particular depending on the body weight, administration route,individual behaviour towards the active compound, nature of theformulation and point of time or interval at which administration takesplace. Thus in some cases it may be sufficient to manage with less thanthe above-mentioned minimum amount, while in other cases the upper limitmentioned must be exceeded. In the case where relatively large amountsare administered, it may be advisable to distribute these into severalindividual doses over the day.

The following embodiment examples illustrate the invention. Theinvention is not limited to the examples.

The percentage data in the following tests and examples are percentagesby weight, unless stated otherwise; parts are parts by weight. Thesolvent ratios, dilution ratios and concentration data of liquid/liquidsolutions in each case relate to the volume.

EXAMPLES

Abbreviations: aq. aqueous solution cat. catalytic d day(s) DCI directchemical ionization (in MS) DMF dimethylformamide DMSO dimethylsulphoxide EI electron impact ionization (in MS) ESI electrosprayionization (in MS) Et ethyl GC-MS gas chromatography-coupled massspectroscopy h hour(s) HPLC high pressure, high performance liquidchromatography conc. concentrated LC-MS liquid chromatography-coupledmass spectroscopy Meth. method min minute(s) MS mass spectroscopy NMRnuclear magnetic resonance spectroscopy R_(t) retention time (in HPLC)RT room temperature TFA trifluoroacetic acid THF tetrahydrofuranLC-MS, GC-MS and HPLC Methods:

Method 1 (LC-MS): instrument: Micromass Platform LCZ with HPLC Agilentseries 1100; column: Thermo Hypersil GOLD 3μ, 20 mm×4 mm; mobile phaseA: 1 l of water+0.5 ml of 50% strength formic acid, mobile phase B: 1 lacetonitrile+0.5 ml of 50% strength formic acid; gradient: 0.0 min 100%A→0.2 min 100% A→2.9 min 30% A→3.1 min 10% A→5.5 min 10% A; oven: 50°C.; flow rate: 0.8 ml/min; UV detection: 210 nm.

Method 2 (LC-MS): instrument type MS: Micromass ZQ; instrument typeHPLC: Waters Alliance 2795; column: Phenomenex Synergi 2μ Hydro-RPMercury 20 mm×4 mm; mobile phase A: 1 l of water+0.5 ml of 50% strengthformic acid, mobile phase B: 1 l acetonitrile+0.5 ml of 50% strengthformic acid; gradient: 0.0 min 90% A→2.5 min 30% A→3.0 min 5% A→4.5 min5% A; flow rate: 0.0 min 1 ml/min ˜2.5 min/3.0 min/4.5 min 2 ml/min;oven: 50° C.; UV detection: 210 nm.

Method 3 (LC-MS): instrument type MS: Micromass ZQ; instrument typeHPLC: Waters Alliance 2795; column: Phenomenex Synergi 2.5μ MAX-RP 100AMercury 20 mm×4 mm; mobile phase A: 1 l of water+0.5 ml of 50% strengthformic acid, mobile phase B: 1 l acetonitrile+0.5 ml of 50% strengthformic acid; gradient: 0.0 min 90% A→0.1 min 90% A→3.0 min 5% A→4.0 min5% A→4.01 min 90% A; flow rate: 2 ml/min; oven: 50° C.; UV detection:210 nm.

Method 4 (LC-MS): instrument: Micromass Quattro Micro MS with HPLCAgilent series 1100; column: Thermo Hypersil GOLD 3μ 20 mm×4 mm; mobilephase A: 1 l of water+0.5 ml of 50% strength formic acid, mobile phaseB: 1 l acetonitrile+0.5 ml of 50% strength formic acid; gradient: 0.0min 100% A→3.0 min 10% A→4.0 min 10% A→4.01 min 100% A (flow rate 2.5ml/min)→5.00 min 100% A; oven: 50° C.; flow rate: 2 ml/min; UVdetection: 210 nm.

Method 5 (LC-MS): instrument: Micromass QuattroPremier with Waters HPLCAcquity; column: Thermo Hypersil GOLD 1.9μ 50 mm×1 mm; mobile phase A: 1l of water+0.5 ml of 50% strength formic acid, mobile phase B: 1 lacetonitrile+0.5 ml of 50% strength formic acid; gradient: 0.0 min 90%A→0.1 min 90% A→1.5 min 10% A→2.2 min 10% A; flow rate: 0.33 ml/min;oven: 50° C.; UV detection: 210 nm.

Method 6 (HPLC): instrument: HP 1100 with DAD detection; column:Kromasil 100 RP-18, 60 mm×2.1 mm, 3.5 μm; mobile phase A: 5 ml ofperchloric acid (70% strength)/litre of water, mobile phase B:acetonitrile; gradient: 0 min 2% B→0.5 min 2% B→4.5 min 90% B→6.5 min90% B→6.7 min 2% B→7.5 min 2% B; flow rate: 0.75 ml/min; columntemperature: 30° C.; UV detection: 210 nm.

Method 7 (GC-MS): instrument: Micromass GCT, GC6890; column: RestekRTX-35, 15 m×200 μm×0.33 μm; constant helium flow: 0.88 ml/min; oven:70° C.; inlet: 250° C.; gradient: 70° C., 30° C./min ˜310° C.(maintained for 3 min).

Method 8 (preparative HPLC): column: Kromasil 100 C18 5 μm, 250 mm×20mm; mobile phase A: Milli-Q water, mobile phase B: aqueous 0.1% strengthtrifluoroacetic acid, mobile phase C: acetonitrile; gradient: 0.0 min76% A, 5% B, 19% C→15 min 4% A, 95% B, 1% C→15.1 min 76% A, 5% B, 19%C→20 min 76% A, 5% B, 19% C; oven: 40° C.; flow rate: 25 ml/min; UVdetection: 210 nm.

Method 9 (preparative HPLC): column: Sunfire C18 5 μm, 19 mm×150 mm;mobile phase A: aqueous 0.2% strength trifluoroacetic acid, mobile phaseB: acetonitrile; gradient: 0.0 min 95% A 8 min 50% A→8.01 min 95% A→12min 95% A; RT; flow rate: 25 ml/min; UV detection: 210 nm.

Method 10 (preparative HPLC): column: Sunfire C18 5 μm, 19 mm×150 mm;mobile phase A: aqueous 0.2% strength trifluoroacetic acid, mobile phaseB: acetonitrile; 0 min 90% A→13 min 90% A; oven: 40° C.; flow rate: 25ml/min; UV detection: 210 nm.

Method 11 (preparative HPLC): column: XBridge C18 5 μm, 19 mm×150 mm;mobile phase A: aqueous 0.2% strength formic acid, mobile phase B:acetonitrile; 0 min 75% A→6 min 75% A; RT; flow rate: 25 ml/min; UVdetection: 210 nm.

Method 12 (preparative HPLC): column: XBridge C18 5 μm, 19 mm×150 mm;mobile phase A: aqueous 0.2% strength formic acid, mobile phase B:acetonitrile; 0 min 93% A→4 min 93% A; RT; flow rate: 25 ml/min; UVdetection: 210 nm.

Method 13 (preparative HPLC): column: XBridge C18 5 μm, 19 mm×150 mm;mobile phase A: aqueous 0.2% strength trifluoroacetic acid, mobile phaseB: acetonitrile; 0 min 90% A→12 min 90% A; oven: 40° C.; flow rate: 25ml/min; UV detection: 210 nm.

Starting Materials Example 1A Ethyl (4-cyano-1H-imidazol-1-yl)acetate

3.3 g (35.3 mmol) of 1H-imidazole-4-carbonitrile [Matthews et al., J.Org. Chem. 1986, 51, 3228-3231] are initially charged in 13.2 ml (11.5g, 35.3 mmol) of 21% strength sodium ethoxide solution in ethanol, and4.3 ml (6.5 g, 38.9 mmol) of ethyl bromoacetate are added. The reactionmixture is stirred at RT for 16 h. For work-up, the precipitated solidis filtered off, the filter residue is washed with ethanol and thefiltrate is concentrated under reduced pressure. Diisopropyl ether isadded to the residue, the mixture is filtered again, the filtrate isonce more concentrated on a rotary evaporator and the residue is driedunder reduced pressure. Yield: 3.8 g (60% of theory)

LC-MS (Method 1): R_(t)=1.17 min; MS (ESIpos): m/z=180 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.12 (s, 1H), 7.88 (s, 1H), 5.06 (s, 2H),4.18 (q, 2H), 1.22 (t, 3H).

Example 2A Ethyl 2-(1H-1,2,3-triazol-1-yl)acetate

129.2 g (5.6 mmol) of sodium are added slowly to 4.0 litres of ethanol.400.0 g (5.6 mol) of 1,2,3-1H-triazole are then added, and 623 ml (938.2g, 5.6 mol) of ethyl bromoacetate are added dropwise at an internaltemperature of 20-25° C. The mixture is stirred at RT for 48 h. Theprecipitated solid is filtered off, the ethanol is removed under reducedpressure and the mixture is filtered again. The residue is taken up inethyl acetate, filtered, again concentrated under reduced pressure andpurified by distillation on a 30 cm column. The product is obtained at abath temperature of 140° C., a head temperature of 60-115° C. and apressure of 1 mbar. Yield: 440.0 g (50% of theory)

HPLC (Method 6): R_(t)=1.58 min;

LC-MS (Method 1): R_(t)=0.71 min; MS (ESIpos): m/z=156 [M+H]⁺.

Example 3A Ethyl 1H-imidazol-1-ylacetate

118.2 g (5.1 mmol) of sodium are added slowly to 2.5 litres of ethanol.350.0 g (5.1 mol) of imidazole and 570 ml (858.6 g, 5.1 mol) of ethylbromoacetate are added dropwise at an internal temperature of 20-25° C.The mixture is stirred at RT for 24 h. The precipitated solid isfiltered off, the ethanol is removed under reduced pressure and themixture is filtered again. The residue is purified by columnchromatography on silica gel (mobile phase ethyl acetate). Yield: 639.0g (81% of theory)

GC-MS (Method 7): R_(t)=4.55 min; MS (ESIpos): m/z=155 [M+H]⁺.

Example 4A Ethyl (4-cyano-1H-1,2,3-triazol-1-yl)acetate

4.1 g (31.9 mmol) of ethyl azidoacetate and 2.8 g (31.9 mmol) of2-chloroacrylonitrile are stirred in 32 ml of water at a bathtemperature of 80° C. for 16 h. After cooling to RT, the solution isacidified with 1 N hydrochloric acid and extracted with ethyl acetate.The organic phase is dried over sodium sulphate, filtered andconcentrated under reduced pressure. 50 ml of ethanol and 10 drops ofconcentrated sulphuric acid are added to the residue, and the mixture isstirred under reflux for 16 h. For work-up, the reaction mixture isconcentrated under reduced pressure, ethyl acetate is added to theresidue, the suspension is washed with semiconcentrated sodiumbicarbonate solution and the organic phase is dried over sodiumsulphate. The solvent is removed completely on a rotary evaporator andthe solid is dried under reduced pressure. Yield: 1.5 g (25% of theory)

LC-MS (Method 3): R_(t)=0.96 min; MS (ESIpos): m/z=181 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=9.06 (s, 1H), 5.57 (s, 2H), 4.19 (q, 2H),1.22 (t, 3H).

Example 5A Ethyl 3-(N,N-dimethylamino)-2-(1H-imidazol-1-yl)acrylate

38.0 g (244.9 mmol) of the compound from Example 3A are stirred in 126ml (108.1 g, 734.7 mmol) of N,N-dimethylformamide diethyl acetal at abath temperature of 90° C. for 16 h. After cooling, the mixture isconcentrated under reduced pressure, the residue is stirred withdiisopropyl ether and the solid is filtered off and finally washed withdiisopropyl ether. Yield: 49.0 g (95% of theory)

LC-MS (Method 2): R_(t)=2.42 min; MS (ESIpos): m/z=211 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=7.52 (s, 1H), 7.49 (s, 1H), 7.05 (s, 1H),6.91 (s, 1H), 4.02 (q, 2H), 2.63 (br. s, 6H), 1.12 (t, 3H).

Example 6A Ethyl3-(N,N-dimethylamino)-2-(4-cyano-1H-imidazol-1-yl)acrylate

3.8 g (21.4 mmol) of the compound from Example 1A and 7.4 ml (6.3 g,42.8 mmol) of N,N-dimethylformamide diethyl acetal are stirred at a bathtemperature of 100° C. for 16 h. For work-up, the cooled reactionsolution is concentrated on a rotary evaporator and the residue is driedunder reduced pressure. Yield: 5.0 g (73% pure, 73% of theory)

LC-MS (Method 1): R_(t)=2.69 min; MS (ESIpos): m/z=235 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.13 (s, 1H), 7.85 (s, 1H), 7.58 (s, 1H),4.03 (q, 2H), 2.69 (br. s, 6H), 1.12 (t, 3H).

Example 7A Ethyl3-(dimethylamino)-2-(4-cyano-1H-1,2,3-triazol-1-yl)acrylate

1.3 g (7.5 mmol) of the compound from Example 4A and 1.4 ml (1.2 g, 8.2mmol) of N,N-dimethylformamide diethyl acetal are stirred at a bathtemperature of 100° C. for 16 h. For work-up, the cooled reactionsolution is concentrated on a rotary evaporator and the residue is driedunder reduced pressure. Yield: 1.5 g (86% of theory)

LC-MS (Method 4): R_(t)=1.55 min; MS (ESIpos): m/z=236 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=9.14 (s, 1H), 7.75 (s, 1H), 4.04 (q, 2H),3.15 (br. s, 3H), 2.18 (br. s, 3H), 1.13 (t, 3h).

Example 8A Ethyl 3-(dimethylamino)-2-(1H-1,2,3-triazol-1-yl)acrylate

44.2 ml (38.0 g, 257.8 mmol) of N,N-dimethylformamide diethyl acetal areadded to 20.0 g (128.9 mmol) of the compound from Example 2A, and themixture is stirred at 100° C. for 16 h. After cooling to RT, thereaction mixture is concentrated under reduced pressure. The residue istriturated with diethyl ether, filtered off and washed with diethylether. Yield: 18.0 g (67% of theory)

LC-MS (Method 4): R_(t)=1.20 min; MS (ESIpos): m/z=211 [M+H]⁺.

¹H-NMR (400 MHz, DMSO-d₆): δ=8.10 (d, 1H), 7.78 (d, 1H), 7.65 (s, 1H),4.03 (q, 2H), 3.06 (br. s, 3H), 2.10 (br. s, 3H), 1.12 (t, 3H).

Example 9A 4-(4-Cyclobutylpiperazin-1-yl)-6-hydrazinopyrimidine

Step a): 4-Chloro-6-(4-cyclobutylpiperazin-1-yl)pyrimidine

1.8 g (8.4 mmol) of 1-cyclobutylpiperazine dihydrochloride (Zaragoza etal., J. Med. Chem. 2004, 47, 2833) are initially charged in 18 ml ofwater, and 2.9 ml (2.1 g, 16.9 mmol) ofN-ethyl-N-(propan-2-yl)propane-2-amine are added. The mixture is stirredat RT for 30 min, and 1.3 g (8.4 mmol) of 4,6-dichloropyrimidine areadded. The reaction mixture is stirred at 115° C. for 1 h and cooled toRT, 25 ml of ethyl acetate are added and the mixture is extracted with25 ml of a saturated aqueous sodium bicarbonate solution. The organicphase is separated off, dried over sodium sulphate, filtered andconcentrated under reduced pressure. The crude product is purified bycolumn chromatography on silica gel (mobile phase:dichloromethane/methanol 100/3). Yield: 1.9 g (89% of theory)

HPLC (Method 6): R_(t)=2.79 min; MS (DCI): m/z=254 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.32 (s, 1H), 6.95 (s, 1H), 3.65-3.58 (m,4H), 2.70 (quintet, 1H), 2.27 (t, 4H), 2.00-1.93 (m, 2H), 1.87-1.75 (m,2H), 1.67-1.55 (m, 2H).

Step b): 4-(4-Cyclobutylpiperazin-1-yl)-6-hydrazinopyrimidine

At RT, 4.4 ml (4.5 g, 89.7 mmol) of hydrazine hydrate are added dropwisewith stirring to a solution of 1.9 g (7.5 mmol) of4-chloro-6-(4-cyclobutylpiperazin-1-yl)pyrimidine in 28 ml of ethanol.The reaction solution is stirred at 80° C. for 16 h. For work-up, themixture is concentrated under reduced pressure, the residue istriturated repeatedly with diethyl ether, and the precipitated solid isfiltered off and dried under reduced pressure. The residue is thenpurified by column chromatography on silica gel (mobile phasedichloromethane/methanol 10/2). Yield: 1.5 g (80% of theory)

LC-MS (Method 6): R_(t)=1.36 min; MS (ESIpos): m/z=249 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=7.92 (s, 1H), 7.63 (s, 1H), 5.90 (NH), 4.09(s, NH2), 3.45 (t, 4H), 2.69 (quintet, 1H), 2.26 (t, 4H), 2.00-1.93 (m,2H), 1.82-1.75 (m, 2H), 1.67-1.60 (m, 2H).

Example 10A 1-(6-Hydrazinylpyrimidin-4-yl)azetidin-3-ol

Step a): 1-(6-Chloropyrimidin-4-yl)azetidin-3-ol

7.3 g (48.7 mmol) of 4,6-dichloropyrimidine are suspended in 140 ml ofwater, and 47 ml 1 N aqueous sodium hydroxide solution are added. 5.3 g(48.7 mmol) of 3-hydroxyazetidine are added, and the reaction mixture isstirred at 90° C. for 3 d. After cooling to RT, the reaction mixture isconcentrated under reduced pressure and reacted further without furtherpurification.

LC-MS (Method 5): R_(t)=0.36 min; MS (ESIpos): m/z=1.87 [M+H]⁺.

Step b): 1-(6-Hydrazinylpyrimidin-4-yl)azetidin-3-ol

At RT, 27.2 ml (27.9 g, 279.1 mmol) of hydrazine hydrate are addeddropwise with stirring to a solution of 10.4 g (55.8 mmol) of1-(6-chloropyrimidin-4-yl)azetidin-3-ol in 100 ml of ethanol. Thereaction solution is stirred at 80° C. for 16 h. For work-up, themixture is concentrated under reduced pressure, and the precipitate isfiltered off and washed twice with in each case 10 ml of ethanol. Yield:2.0 g (19% of theory)

LC-MS (Method 1): R_(t)=2.06 min; MS (ESIpos): m/z=194 [M+H]⁺.

Example 11A 4-Chloro-6-hydrazinopyrimidine

At RT, 11.8 ml (12.1 g, 241.6 mmol) of hydrazine hydrate are addeddropwise with stirring to a solution of 20.0 g (134.3 mmol) of4,6-dichloropyrimidine in 300 ml of ethanol. If the solution becomesturbid during the metered addition of the hydrazine hydrate, furtherethanol (about 400 ml) is added. The reaction solution is stirred at RTfor 12 h. For work-up, the precipitated solid is filtered off, thefilter residue is washed twice with in each case 150 ml of water andtwice with in each case 100 ml of diethyl ether and the product is driedunder reduced pressure. The concentrated mother liquor gives a furthercrystalline product fraction. Yield: 16.8 g (87% of theory)

LC-MS (Method 1): R_(t)=1.17 min; MS (ESIpos): m/z=145 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.81 (s, 1H), 8.17 (br. s, 1H), 6.75 (s,1H), 4.48 (br. s, 2H).

Example 12A2-(6-Chloropyrimidin-4-yl)-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-onehydrochloride

10.0 g (47.7 mmol) of the compound from Example 8A and 8.3 g (57.1 mmol)of the compound from Example 11A are initially charged in 100 ml ofethanol, and 1.5 ml (2.2 g, 19.0 mmol) of trifluoroacetic acid areadded. The mixture is stirred under reflux for 12 h. An excess of a 4 Msolution of hydrogen chloride in dioxane is then added to the cooledreaction mixture, the mixture is stirred for about 1 h, the precipitatedcrystals are filtered off and the filter residue is washed with dioxaneand ethanol. The intermediate obtained in this manner is dissolved in150 ml of ethanol, 50 ml of a 25% strength methanolic sodium methoxidesolution are added and the mixture is stirred at RT for 2 h. Thereaction mixture is then adjusted with 1 N hydrochloric acid to pH=5 andstirred at RT for a further 2 h, the solid is filtered off, the filterresidue is washed with ethanol and the product is dried under reducedpressure. Yield: 7.0 g (49% of theory)

LC-MS (Method 5): R_(t)=1.20 min; MS (ESIpos): m/z=264 [M+H]⁺.

Example 13A2-(6-Chloropyrimidin-4-yl)-4-(1H-imidazol-1-yl)-1,2-dihydro-3H-pyrazol-3-onehydrochloride

10.0 g (47.8 mmol) of the compound from Example 5A and 8.3 g (57.3 mmol)of the compound from Example 11A are initially charged in 100 ml ofethanol, and 1.5 ml (2.2 g, 19.0 mmol) of trifluoroacetic acid areadded. The mixture is stirred under reflux for 12 h. The precipitatedcrystals are filtered off, the filter residue is washed with ethanol andthe intermediate is dried under reduced pressure overnight. Theintermediate is then suspended in 20 ml of methanol, 100 ml of a 4 Msolution of hydrogen chloride in dioxane are added and the mixture isstirred at RT for 1 h. The solid is filtered off, the filter residue iswashed with dioxane, ethyl acetate and diisopropyl ether and the productis dried under reduced pressure. Yield: 4.6 g (32% of theory)

HPLC (Method 6): R_(t)=2.81 min; MS (ESIpos): m/z=263 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=9.46 (s, 1H), 8.96 (s, 1H), 8.56 (s, 1H),8.51 (d, 1H), 8.07-8.04 (m, 1H), 7.85-7.82 (m, 1H).

Example 14A 4-(6-Hydrazinylpyrimidin-4-yl)-1,4-oxazepane

Step a): 4-(6-Chloropyrimidin-4-yl)-1,4-oxazepane

A mixture of 3.0 g (20.1 mmol) of 4,6-dichloropyrimidine, 2.8 g (20.1mmol) of 1,4-oxazepane hydrochloride and 6.4 g (60.4 mmol) of sodiumcarbonate in 45 ml of water is stirred under reflux for 16 h. Aftercooling to RT, the reaction mixture is extracted with ethyl acetate. Theorganic phase is washed with saturated sodium chloride solution, driedover sodium sulphate and filtered. Under reduced pressure, the filtrateis concentrated to dryness. The product is obtained as an oil. Yield:3.9 g (86% of theory)

LC-MS (Method 4): R_(t)=1.32 min; MS (ESIpos): m/z=214 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.33 (s, 1H), 6.86 (s, 1H), 3.99-3.52 (m,8H), 1.84 (m, 2H).

Step b): 4-(6-Hydrazinylpyrimidin-4-yl)-1,4-oxazepane

At RT, 8.8 ml (9.0 g, 180.2 mmol) of hydrazine hydrate are addeddropwise with stirring to a solution of 3.9 g (18.0 mmol) of4-(6-chloropyrimidin-4-yl)-1,4-oxazepane in 25 ml of ethanol. After 16 hof stirring at 80° C., the reaction solution is concentrated underreduced pressure. The residue is triturated with cold ethanol, theprecipitated solid is filtered off and the filter residue is washed with25 ml of diethyl ether. The product is dried under reduced pressure.Yield: 1.4 g (36% of theory)

HPLC (Method 11): R_(t)=2.48 min; MS (ESIpos): m/z=210 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=7.91 (s, 1H), 7.56 (br. s, 1H), 5.81 (s,1H), 4.12 (br. s, 2H), 3.75-3.55 (m, 8H), 1.85 (quintet, 2H).

Example 12-[6-(4-Cyclobutylpiperazin-1-yl)pyrimidin-4-yl]-4-(1H-imidazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one

16 μl (23 mg, 0.2 mmol) of trifluoroacetic acid are added to a mixtureof 211 mg (1.0 mmol) of the compound from Example 5A and 250 mg (1.0mmol) of the compound from Example 9A in 4 ml of ethyl acetate, and themixture is stirred at 100° C. for 20 h. The reaction mixture isconcentrated under reduced pressure, more ethyl acetate andtrifluoroacetic acid (the same amounts as above) are added and themixture is stirred at 100° C. for 20 h. The reaction mixture is cooledto RT, and the precipitated solid is filtered off and washed withdiethyl ether. The residue is pre-purified by column chromatography onsilica gel (mobile phase dichloromethane/methanol/ammonia 10/2/0.2) andpurified by preparative HPLC (RP18 column; mobile phase:acetonitrile/water gradient). Yield: 137 mg (36% of theory)

HPLC (Method 6): R_(t)=2.73 min; MS (ESIpos): m/z=367 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.40 (s, 1H), 8.12 (s, 1H), 7.90 (s, 1H),7.74 (s, 1H), 7.50 (s, 1H), 7.07 (s, 1H), 3.65-3.58 (m, 4H), 2.77(quintet, 1H), 2.38-2.35 (m, 4H), 2.01-1.96 (m, 2H), 1.89-1.79 (m, 2H),1.67-1.62 (m, 2H).

Example 22-[6-(4-Cyclobutylpiperazin-1-yl)pyrimidin-4-yl]-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one

16 μl (23 mg, 0.2 mmol) of trifluoroacetic acid are added to a mixtureof 211 mg (1.0 mmol) of the compound from Example 8A and 250 mg (1.0mmol) of the compound from Example 9A in 4 ml of ethyl acetate, and themixture is stirred at 100° C. for 20 h. The reaction mixture isconcentrated under reduced pressure, more ethyl acetate andtrifluoroacetic acid (the same amounts as above) are added and themixture is stirred at 100° C. for 3 d. The reaction mixture is cooled toRT, and the precipitated solid is filtered off and washed with diethylether. The residue is suspended in 2 ml of water, dissolved by addingaqueous 1 N sodium hydroxide solution (pH=9-10) and purified bypreparative HPLC (RP18 column; mobile phase: acetonitrile/watergradient). Yield: 195 mg (51% of theory)

HPLC (Method 6): R_(t)=2.90 min; MS (ESIpos): m/z=368 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.41 (s, 1H), 8.40 (s, 1H), 7.88 (s, 1H),7.82 (s, 1H), 7.75 (s, 1H), 3.70-3.65 (m, 4H), 3.03-2.97 (m, 1H),2.60-2.57 (m, 4H), 2.06-2.02 (m, 2H), 1.98-1.90 (m, 2H), 1.70-1.64 (m,2H).

Example 31-{2-[6-(4-Cyclobutylpiperazin-1-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-imidazole-4-carbonitrile

16 μl (23 mg, 0.2 mmol) of trifluoroacetic acid are added to a mixtureof 236 mg (1.0 mmol) of the compound from Example 6A and 250 mg (1.0mmol) of the compound from Example 9A in 4 ml of ethyl acetate, and themixture is stirred at 100° C. for 20 h. The reaction mixture isconcentrated under reduced pressure, more ethyl acetate andtrifluoroacetic acid (the same amounts as above) are added and themixture is stirred at 100° C. for 3 d. The reaction mixture is cooled toRT, and the precipitated solid is filtered off and washed with diethylether. The residue is suspended in 2 ml of water, dissolved by addingaqueous 1 N sodium hydroxide solution (pH=9-10) and purified bypreparative HPLC (RP18 column; mobile phase: acetonitrile/watergradient). Yield: 219 mg (56% of theory)

HPLC (Method 6): R_(t)=3.10 min; MS (ESIpos): m/z=392 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.38 (d, 1H), 8.37 (d, 1H), 8.17 (d, 1H),7.82 (s, 1H), 7.80 (s, 1H), 3.68-3.62 (m, 4H), 3.38-3.30 (m, 4H),2.96-2.91 (m, 1H), 2.06-2.00 (m, 2H), 1.95-1.85 (m, 2H), 1.70-1.63 (m,2H).

Example 41-{2-[6-(3-Hydroxyazetidin-1-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-imidazole-4-carbonitrile

46 μl (68 mg, 0.6 mmol) of trifluoroacetic acid are added to a mixtureof 700 mg (3.0 mmol) of the compound from Example 6A and 541 mg (3.0mmol) of the compound from Example 10A in 10 ml of ethyl acetate, andthe mixture is stirred at 100° C. for 10 h. The reaction mixture isconcentrated under reduced pressure and taken up in 5 ml of ethanol, andthe precipitate is filtered off. The solid is suspended in 10 ml ofwater, and 1 N aqueous sodium hydroxide solution (pH=9) is added untilthe solid is dissolved. The pH is then adjusted to 7 using 1 Nhydrochloric acid, the mixture is concentrated to a volume of about 5 mland the precipitate formed is filtered off. The residue is washed withwater and diisopropyl ether and chromatographed by preparative HPLC(Method 8). The solid is then suspended in 10 ml of water, and 1 Naqueous sodium hydroxide solution (pH=9) is added until the solid isdissolved. The pH is then adjusted to 7 using 1 N hydrochloric acid, themixture is concentrated to a volume of about 2.5 ml and the precipitateformed is filtered off. The filtrate is concentrated to about 2 ml andfiltered again. Both residues are combined, washed with water and ethylacetate and dried under reduced pressure. Yield: 78 mg (8% of theory)

HPLC (Method 6): R_(t)=2.90 min; MS (ESIpos): m/z=325 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.35 (s, 1H), 8.29 (s, 1H), 8.17 (s, 1H),7.66 (s, 1H), 7.34 (s, 1H), 5.80-5.75 (m, 1H), 4.63-4.58 (m, 1H),4.24-4.20 (m, 2H), 3.75-3.73 (m, 2H).

Example 51-{6-[4-(1H-Imidazol-1-yl)-5-oxo-2,5-dihydro-1H-pyrazol-1-yl]pyrimidin-4-yl}azetidine-3-carboxylicacid

46 mg (0.3 mmol) of azetidine-3-carboxylic acid hydrochloride areinitially charged in a mixture of 1 ml of water and 0.3 ml of ethanol.100 mg (0.3 mmol) of the compound from Example 13A are added, and themixture is stirred at 100° C. for 1 h. The reaction mixture is thenadjusted to pH=7 using aqueous 1 N sodium hydroxide solution and stirredat 100° C. for 16 h. Once more, the mixture is adjusted to pH=7 usingaqueous 1 N sodium hydroxide solution and reacted in a single modemicrowave (Emrys Optimizer) at 150° C. for 1 h. The reaction mixture isconcentrated under reduced pressure and purified by preparative HPLC(RP18 column; mobile phase: acetonitrile/water gradient). Yield: 23 mg(21% of theory)

LC-MS (Method 8): R_(t)=0.86 min; MS (ESIpos): m/z=328 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.41 (s, 1H), 7.79 (s, 1H), 7.47 (s, 1H),7.36 (s, 1H), 7.31 (s, 1H), 6.89 (s, 1H), 4.09 (t, 2H), 3.99 (t, 2H).

Example 61-{6-[5-Oxo-4-(1H-1,2,3-triazol-1-yl)-2,5-dihydro-1H-pyrazol-1-yl]pyrimidin-4-yl}azetidine-3-carboxylicacid

55 mg (0.4 mmol) of azetidine-3-carboxylic acid hydrochloride areinitially charged in 2 ml of water. 100 mg (0.3 mmol) of the compoundfrom Example 12A are added, and the pH is adjusted to 7 using aqueous 1N sodium hydroxide solution. The mixture is reacted in a single modemicrowave (Emrys Optimizer) at 150° C. for 1 h. The reaction mixture isconcentrated under reduced pressure and purified by preparative HPLC(RP18 column; mobile phase: acetonitrile/water gradient). Yield: 15 mg(13% of theory)

HPLC (Method 6): R_(t)=2.81 min; MS (ESIpos): m/z=329 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.42 (s, 1H), 8.26 (s, 1H), 7.70 (s, 1H),7.67 (s, 1H), 7.34 (s, 1H), 4.05-3.96 (m, 2H and 2H), 3.13-3.07 (m, 1H).

Example 74-(1H-Imidazol-1-yl)-2-[6-(3-methylazetidin-1-yl)pyrimidin-4-yl]-1,2-dihydro-3H-pyrazol-3-one

43 mg (0.4 mmol) of 3-methylazetidine hydrochloride, 100 mg (0.3 mmol)of the compound from Example 13A and 174 μl (130 mg, 1.0 mmol) ofN-ethyl-N-(propan-2-yl)propane-2-amine are suspended in 2 ml oftetrahydrofuran and reacted in a single mode microwave (Emrys Optimizer)at 120° C. for 4.5 h. The reaction mixture is concentrated under reducedpressure, taken up in water with addition of aqueous 1 N sodiumhydroxide solution (pH=9-10) and purified by preparative HPLC (RP18column; mobile phase: acetonitrile/water gradient). Yield: 30 mg (30% oftheory)

HPLC (Method 6): R_(t)=3.07 min; MS (ESIpos): m/z=298 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.25 (s, 1H), 7.81 (s, 1H), 7.46 (s, 1H),7.39 (s, 1H), 7.31 (s, 1H), 6.88 (s, 1H), 4.10 (t, 2H), 3.54 (dd, 2H),2.86-2.75 (m, 1H), 1.25 (d, 3H).

Example 82-[6-(3-Methylazetidin-1-yl)pyrimidin-4-yl]-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one

43 mg (0.4 mmol) of 3-methylazetidine hydrochloride, 100 mg (0.3 mmol)of the compound from Example 12A and 174 μl (130 mg, 1.0 mmol) ofN-ethyl-N-(propan-2-yl)propane-2-amine are suspended in 2 ml oftetrahydrofuran and reacted in a single mode microwave (Emrys Optimizer)at 120° C. for 1.5 h. The reaction mixture is concentrated under reducedpressure, taken up in water with addition of aqueous 1 N sodiumhydroxide solution (pH=9-10) and purified by preparative HPLC (RP18column; mobile phase: acetonitrile/water gradient). Yield: 25 mg (25% oftheory)

HPLC (Method 6): R_(t)=3.00 min; MS (ESIpos): m*z=299 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.41 (s, 1H), 8.27 (s, 1H), 7.70 (s, 1H),7.67 (s, 1H), 7.41 (s, 1H), 4.11 (t, 2H), 3.56 (dd, 2H), 2.86-2.78 (m,1H), 1.25 (d, 3H).

Example 92-{6-[3-(Dimethylamino)azetidin-1-yl]pyrimidin-4-yl}-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-onedihydrochloride

271 mg (1.5 mmol) of N,N-dimethylazetidine-3-amine dihydrochloride, 400mg (1.5 mmol) of the compound from Example 12A and 847 mg (6.1 mmol) ofpotassium carbonate are suspended in 8 ml of N,N-dimethylformamide andstirred at 100° C. for 16 h. The reaction mixture is concentrated underreduced pressure and purified by preparative HPLC (RP18 column; mobilephase: acetonitrile/water gradient with addition of 0.1% strengthtrifluoroacetic acid). Further purification is carried out bypreparative HPLC (RP18 column; mobile phase: acetonitrile/water gradientwith addition of 0.1% strength formic acid). 2 ml of 1 N hydrochloricacid is added to the product-containing fractions, and the mixture isstirred at RT for 1 h. The solid is filtered off and dried under highvacuum. Yield: 62 mg (17% of theory)

LC-MS (Method 5): R_(t)=0.19 min; MS (ESIpos): m/z=328 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.42 (s, 1H), 8.28 (s, 1H), 7.69 (s, 1H),7.66 (s, 1H), 7.52 (s, 1H), 4.02 (t, 2H), 3.76 (dd, 2H), 3.24-3.18 (m,1H), 2.12 (s, 6H).

Example 102-[6-(4,4-Difluoropiperidin-1-yl)pyrimidin-4-yl]-4-(1H-imidazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one

100 mg (0.3 mmol) of the compound from Example 13A, 63 mg (0.4 mmol) of4,4-difluoropiperidine hydrochloride and 116 μl (86 mg, 0.7 mmol) ofN-ethyl-N-(propan-2-yl)propane-2-amine are initially charged in 2 ml oftetrahydrofuran and reacted in a single mode microwave (Emrys Optimizer)at 120° C. for 2.5 h. After concentration under reduced pressure, theresidue is taken up in acetonitrile/water and purified by preparativeHPLC (RP18 column; mobile phase: acetonitrile/water gradient). Yield: 82mg (71% of theory)

HPLC (Method 6): R_(t)=3.37 min; MS (ESIpos): m/z=348 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.49 (s, 1H), 8.38 (s, 1H), 8.15 (s, 1H),7.76 (s, 1H), 7.64 (s, 1H), 7.22 (s, 1H), 3.80 (t, 4H), 2.06 (heptet,4H).

Example 112-[6-(4,4-Difluoropiperidin-1-yl)pyrimidin-4-yl]-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one

250 mg (0.8 mmol) of the compound from Example 12A, 158 mg (1.0 mmol) of4,4-difluoropiperidine hydrochloride and 435 μl (323 mg, 2.5 mmol) ofN-ethyl-N-(propan-2-yl)propane-2-amine are initially charged in 5 ml oftetrahydrofuran and reacted in a single mode microwave (Emrys Optimizer)at 120° C. for 30 min. After pre-purification by preparative HPLC (RP18column; mobile phase: acetonitrile/water gradient), the product isadditionally purified by column chromatography on silica gel (mobilephase dichloromethane/methanol, 10/1). Yield: 29 mg (10% of theory)

HPLC (Method 6): R_(t)=3.49 min; MS (ESIpos): m/z=349 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.56 (s, 1H), 8.39 (s, 1H), 8.30 (s, 1H),7.87 (s, 1H), 7.57 (s, 1H), 3.91-3.81 (m, 4H), 2.09 (heptet, 4H).

Example 121-{2-[6-(4,4-Difluoropiperidin-1-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-imidazole-4-carbonitrile

A mixture of 200 mg (1.4 mmol) of the compound from Example 11A, 262 mg(1.7 mmol) of 4,4-difluoropiperidine hydrochloride and 289 μl (215 mg,1.7 mmol) of N-ethyl-N-(propan-2-yl)propane-2-amine in 3 ml of water isstirred at 100° C. for 16 h. Following the addition of 53 μl (79 mg, 0.7mmol) of trifluoroacetic acid and 324 mg (1.4 mmol) of the compound fromExample 6A, the reaction mixture is stirred at 100° C. for 16 h. Theprecipitated solid is filtered off and washed first with water and thenwith diethyl ether. The product is dried under reduced pressure. Yield:111 mg (21% of theory)

LC-MS (Method 4): R_(t)=1.69 min; MS (ESIpos): m/z=373 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.55 (s, 1H), 8.44 (d, 1H), 8.33 (s, 1H),8.22 (d, 1H), 7.60 (br. s, 1H), 3.84 (br. s, 4H), 2.09 (heptet, 4H).

Example 131-{2-[6-(4,4-Difluoropiperidin-1-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-1,2,3-triazole-4-carbonitrile

A mixture of 200 mg (1.4 mmol) of the compound from Example 11A, 262 mg(1.7 mmol) of 4,4-difluoropiperidine hydrochloride and 289 μl (215 mg,1.7 mmol) of N-ethyl-N-(propan-2-yl)propane-2-amine in 3 ml of water isstirred at 100° C. for 16 h. Following the addition of 53 μl (79 mg, 0.7mmol) of trifluoroacetic acid and 325 mg (1.4 mmol) of the compound fromExample 7A, the reaction mixture is stirred at 100° C. for 16 h. Theprecipitated solid is filtered off and washed first with water and thenwith diethyl ether. The product is dried under reduced pressure. Yield:34 mg (7% of theory)

LC-MS (Method 4): R_(t)=1.77 min; MS (ESIpos): m/z=374 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=9.24 (s, 1H), 8.59 (s, 1H), 8.27 (s, 1H),7.54 (s, 1H), 3.90 (br. s, 4H), 2.12 (heptet, 4H).

Example 142-[6-(3,3-Difluoropyrrolidin-1-yl)pyrimidin-4-yl]-4-(1H-imidazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one

100 mg (0.3 mmol) of the compound from Example 13A, 58 mg (0.4 mmol) of3,3-difluoropyrrolidine hydrochloride and 175 μl (130 mg, 1.0 mmol) ofN-ethyl-N-(propan-2-yl)propane-2-amine are initially charged in 2 ml oftetrahydrofuran and reacted in a single mode microwave (Emrys Optimizer)at 120° C. for 1 h. After concentration under reduced pressure, theresidue is taken up in acetonitrile/water and purified by preparativeHPLC (RP18 column; mobile phase: acetonitrile/water gradient). Yield:111 mg (99% of theory)

HPLC (Method 6): R_(t)=3.18 min; MS (ESIpos): m/z=334 [M+H]⁺;

¹H-NMR (400 MHz, methanol-d₄): δ=8.41 (s, 1H), 7.85 (s, 1H), 7.62 (s,1H), 7.56 (s, 1H), 7.29 (s, 1H), 7.03 (s, 1H), 3.91 (t, 2H), 3.76 (t,2H), 2.55 (heptet, 2H).

Example 152-[6-(3,3-Difluoropyrrolidin-1-yl)pyrimidin-4-yl]-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one

100 mg (0.8 mmol) of the compound from Example 12A, 57 mg (0.4 mmol) of3,3-difluoropyrrolidine hydrochloride and 174 μl (129 mg, 1.0 mmol) ofN-ethyl-N-(propan-2-yl)propane-2-amine were initially charged in 2 ml oftetrahydrofuran and reacted in a single mode microwave (Emrys Optimizer)at 120° C. for 30 min. After concentration under reduced pressure, theresidue is taken up in acetonitrile/water and purified by preparativeHPLC (RP18 column; mobile phase: acetonitrile/water gradient). Yield: 13mg (12% of theory)

HPLC (Method 6): R_(t)=3.33 min; MS (ESIpos): m/z=335 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.42 (s, 1H), 8.35 (s, 1H), 7.72-7.66 (m,3H), 3.87 (t, 2H), 3.65 (t, 2H), 2.64-2.50 (m, partially beneath theDMSO signal, 2H).

Example 161-{2-[6-(3,3-Difluoropyrrolidin-1-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-imidazole-4-carbonitrile

A mixture of 200 mg (1.4 mmol) of the compound from Example 11A, 238 mg(1.7 mmol) of 3,3-difluoropyrrolidine hydrochloride and 289 μl (215 mg,1.7 mmol) of N-ethyl-N-(propan-2-yl)propane-2-amine in 3 ml of water isstirred at 100° C. for 16 h. Following the addition of 53 μl (79 mg, 0.7mmol) of trifluoroacetic acid and 324 mg (1.4 mmol) of the compound fromExample 6A, the reaction mixture is stirred at 100° C. for 16 h. 1 ml of1 N hydrochloric acid is added to the reaction mixture. The resultingprecipitated hydrochloride of the crude product is filtered off, washedwith diethyl ether and dried. The crude product still contains unreactedstarting material (compound from Example 11A). Accordingly, the crudeproduct is reacted with 108 μl (80 mg, 0.6 mmol) ofN-ethyl-N-(propan-2-yl)propane-2-amine, 10 mg (0.1 mmol) of3,3-difluoropyrrolidine hydrochloride and 2 ml of water in a single modemicrowave (Emrys Optimizer) at 170° C. for 15 min. The reaction solutionpurified by preparative HPLC (RP18 column; mobile phase:acetonitrile/water gradient). Yield: 30 mg (6% of theory)

LC-MS (Method 4): R_(t)=1.58 min; MS (ESIpos): m/z=359 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.55 (s, 1H), 8.44 (d, 1H), 8.33 (s, 1H),8.21 (d, 1H), 7.26 (br. s, 1H), 3.99 (t, 2H), 3.75 (br. s, 2H),2.66-2.54 (m, partially beneath the DMSO signal, 2H).

Example 171-{2-[6-(3,3-Difluoropyrrolidin-1-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-1,2,3-triazole-4-carbonitrile

A mixture of 200 mg (1.4 mmol) of the compound from Example 11A, 238 mg(1.7 mmol) of 3,3-difluoropyrrolidine hydrochloride and 289 μl (215 mg,1.7 mmol) of N-ethyl-N-(propan-2-yl)propane-2-amine in 3 ml of water isstirred at 100° C. for 16 h. Following the addition of 53 μl (79 mg, 0.7mmol) of trifluoroacetic acid and 325 mg (1.4 mmol) of the compound fromExample 7A, the reaction mixture is stirred at 100° C. for 16 h. Theprecipitated solid is filtered off and washed first with water and thenwith diethyl ether. The product is dried under reduced pressure. Yield:137 mg (27% of theory)

LC-MS (Method 4): R_(t)=1.66 min; MS (ESIpos): m/z=360 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=9.25 (s, 1H), 8.61 (s, 1H), 8.29 (s, 1H),7.21 (br. s, 1H), 4.06 (t, 2H), 3.82 (br. s, 2H), 2.69-2.55 (m,partially beneath the DMSO signal, 2H).

Example 182-[6-(3,3-Difluoroazetidin-1-yl)pyrimidin-4-yl]-4-(1H-imidazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one

100 mg (0.3 mmol) of the compound from Example 13A, 52 mg (0.4 mmol) of3,3-difluoroazetidine hydrochloride and 175 μl (130 mg, 1.0 mmol) ofN-ethyl-N-(propan-2-yl)propane-2-amine are initially charged in 2 ml oftetrahydrofuran and reacted in a single mode microwave (Emrys Optimizer)at 120° C. for 3 h. The precipitated solid is filtered off and thefiltrate is concentrated under reduced pressure. The residue is taken upin acetonitrile/water and purified by preparative HPLC (RP18 column;mobile phase: acetonitrile/water gradient). Yield: 36 mg (32% of theory)

HPLC (Method 6): R_(t)=3.00 min; MS (ESIpos): m/z=320 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.42 (s, 1H), 8.16 (s, 1H), 7.79 (s, 1H),7.56 (s, 1H), 7.50 (s, 1H), 7.07 (s, 1H), 4.49 (t, 4H).

Example 192-[6-(3,3-Difluoroazetidin-1-yl)pyrimidin-4-yl]-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one

100 mg (0.3 mmol) of the compound from Example 12A, 52 mg (0.4 mmol) of3,3-difluoroazetidine hydrochloride and 174 μl (130 mg, 1.0 mmol) ofN-ethyl-N-(propan-2-yl)propane-2-amine are initially charged in 2 ml oftetrahydrofuran and reacted in a single mode microwave (Emrys Optimizer)at 120° C. for 30 min. After concentration under reduced pressure, theresidue is taken up in acetonitrile/water and purified by preparativeHPLC (RP18 column; mobile phase: acetonitrile/water gradient). Yield: 36mg (34% of theory)

HPLC (Method 6): R_(t)=3.20 min; MS (ESIpos): m/z=321 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.42 (s, 1H), 8.39 (s, 1H), 7.71 (s, 2H),7.62 (s, 1H), 4.46 (t, 4H).

Example 201-{2-[6-(3,3-Difluoroazetidin-1-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-imidazole-4-carbonitrile

A mixture of 120 mg (0.8 mmol) of the compound from Example 11A, 129 mg(1.0 mmol) of 3,3-difluoroazetidine hydrochloride and 174 μl (129 mg,1.0 mmol) of N-ethyl-N-(propan-2-yl)propane-2-amine in 3 ml of water isstirred at 100° C. for 16 h. Following the addition of 32 μl (47 mg, 0.4mmol) of trifluoroacetic acid and 194 mg (0.8 mmol) of the compound fromExample 6A, the reaction mixture is stirred at 100° C. for 16 h. Theprecipitated solid is filtered off and washed first with water and thenwith diethyl ether. The product is dried under reduced pressure. Yield:32 mg (11% of theory)

LC-MS (Method 4): R_(t)=1.48 min; MS (ESIpos): m/z=345 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.80-8.08 (m, 4H), 7.25 (s, 1H), 4.61 (br.s, 4H).

Example 211-{2-[6-(3,3-Difluoroazetidin-1-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-1,2,3-triazole-4-carbonitrile

A mixture of 120 mg (0.8 mmol) of the compound from Example 11A, 129 mg(1.0 mmol) of 3,3-difluoroazetidine hydrochloride and 174 μl (129 mg,1.0 mmol) of N-ethyl-N-(propan-2-yl)propane-2-amine in 3 ml of water isstirred at 100° C. for 16 h. Following the addition of 32 μl (47 mg, 0.4mmol) of trifluoroacetic acid and 194 mg (0.8 mmol) of the compound fromExample 7A, the reaction mixture is stirred at 100° C. for 16 h. Theprecipitated solid is filtered off and washed first with water and thenwith diethyl ether. The product is dried under reduced pressure. Yield:51 mg (18% of theory)

LC-MS (Method 4): R_(t)=1.56 min; MS (ESIpos): m/z=346 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=9.26 (s, 1H), 8.60 (s, 1H), 8.36 (s, 1H),7.18 (s, 1H), 4.69 (t, 4H).

Example 224-(1H-Imidazol-1-yl)-2-[6-(1,4-oxazepan-4-yl)pyrimidin-4-yl]-1,2-dihydro-3H-pyrazol-3-onehydrochloride

A mixture of 200 mg (1.0 mmol) of the compound from Example 5A, 200 mg(1.0 mmol) of the compound from Example 14A and 37 μl (54 mg, 0.5 mmol)of trifluoroacetic acid in 3 ml of water is stirred at 100° C. for 16 h.After pre-purification by preparative HPLC (RP18 column; mobile phase:acetonitrile/water gradient), the crude product is stirred with 1 ml of1 N hydrochloric acid, filtered off and washed with diethyl ether. Theproduct is dried under reduced pressure. Yield: 21 mg (6% of theory)

LC-MS (Method 4): R_(t)=0.95 min; MS (ESIpos): m/z=364 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=9.46 (s, 1H), 8.56 (s, 1H), 8.30 (s, 1H),8.06 (t, 1H), 7.83 (t, 1H), 7.50-7.33 (m, 1H), 4.05 (br. s, 2H),3.91-3.70 (m, 4H), 3.68 (t, 2H), 2.04-1.73 (m, 2H).

Example 232-[6-(1,4-Oxazepan-4-yl)pyrimidin-4-yl]-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-onehydrochloride

A mixture of 400 mg (1.3 mmol) of the compound from Example 12A and 220mg (1.6 mmol) of 1,4-oxazepane hydrochloride in 4 ml of propan-2-ol isreacted in a single mode microwave (Emrys Optimizer) at 115° C. for 30min. 232 μl (172 mg, 1.3 mmol) of N-ethyl-N-(propan-2-yl)propane-2-amineare added, and the reaction mixture is reacted in a single modemicrowave (Emrys Optimizer) at 115° C. for 20 min. After concentrationunder reduced pressure, the residue is taken up inacetonitrile/water/trifluoroacetic acid and chromatographed bypreparative HPLC (Method 9). The trifluoroacetate salt obtained from theHPLC separation is lyophilized and converted into the hydrochlorideusing 2 ml of 1 N hydrochloric acid. Yield: 7 mg (1% of theory)

LC-MS (Method 4): R_(t)=1.20 min; MS (ESIpos): m/z=329 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.54 (s, 1H), 8.38 (s, 1H), 8.25 (s, 1H),7.86 (d, 1H), 7.35 (br. s, 1H), 4.14-3.69 (m, 6H), 3.67 (t, 2H),2.03-1.77 (m, 2H).

Example 242-[6-(1,4-Oxazepan-4-yl)pyrimidin-4-yl]-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one

A mixture of 500 mg (1.7 mmol) of the compound from Example 12A, 688 mg(5.0 mmol) of 1,4-oxazepane hydrochloride and 1.4 ml (1077 mg, 8.3 mmol)of N-ethyl-N-(propan-2-yl)propane-2-amine is reacted in 10 ml oftetrahydrofuran/ethanol (1/1) in a single mode microwave (EmrysOptimizer) at 140° C. for 30 min. After pre-purification by preparativeHPLC (RP18 column; mobile phase: acetonitrile/water gradient), the crudeproduct is taken up in acetonitrile/trifluoroacetic acid andchromatographed by preparative HPLC (Method 10). The trifluoroacetatesalt obtained from the HPLC separation is lyophilized and adjusted topH=7-8 using 1 N aqueous sodium hydroxide solution. The product isisolated by preparative HPLC (RP18 column; mobile phase:acetonitrile/water gradient). Yield: 176 mg (31% of theory)

LC-MS (Method 4): R_(t)=1.19 min; MS (ESIpos): m/z=329 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.41 (d, 1H), 8.33 (d, 1H), 7.78 (br. s,1H), 7.72-7.70 (m, 2H), 3.91-3.65 (m, 6H), 3.62 (t, 2H), 1.95-1.83 (m,2H).

Example 251-{2-[6-(1,4-Oxazepan-4-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-imidazole-4-carbonitrile

A mixture of 200 mg (0.9 mmol) of the compound from Example 6A, 178 mg(0.9 mmol) of the compound from Example 14A and 33 μl (49 mg, 0.4 mmol)of trifluoroacetic acid in 3 ml of water is stirred at 100° C. for 16 h.The precipitated solid is filtered off and washed first with water andthen with diethyl ether. The product is dried under reduced pressure.Yield: 120 mg (40% of theory)

HPLC (Method 6): R_(t)=3.27 min; MS (ESIpos): m/z=353 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.52 (s, 1H), 8.42 (d, 1H), 8.25 (s, 1H),8.19 (d, 1H), 7.39 (br. s, 1H), 4.14-3.70 (m, 6H), 3.66 (t, 2H),2.05-1.68 (m, 2H).

Example 261-{2-[6-(1,4-Oxazepan-4-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-1,2,3-triazole-4-carbonitrile

A mixture of 200 mg (0.9 mmol) of the compound from Example 7A, 178 mg(0.9 mmol) of the compound from Example 14A and 33 μl (49 mg, 0.4 mmol)of trifluoroacetic acid in 3 ml of water is stirred at 100° C. for 16 h.The precipitated solid is filtered off and washed first with water andthen with diethyl ether. The product is dried under reduced pressure.Yield: 80 mg (27% of theory)

LC-MS (Method 4): R_(t)=1.40 min; MS (ESIpos): m/z=354 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=9.22 (s, 1H), 8.56 (s, 1H), 8.21 (s, 1H),7.45-7.26 (m, 1H), 4.06 (br. s, 2H), 3.92-3.71 (m, 4H), 3.68 (t, 2H),2.06-1.74 (m, 2H).

Example 272-[6-(1,2-Oxazinan-2-yl)pyrimidin-4-yl]-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one

A mixture of 200 mg (0.7 mmol) of the compound from Example 12A, 99 mg(0.8 mmol) of 1,2-oxazinane hydrochloride [Bhat et al., J. Chem. Soc.Perkin Trans. 2 2000, 7, 1435-1446] and 348 μl (258 mg, 2.0 mmol) ofN-ethyl-N-(propan-2-yl)propane-2-amine in 4 ml of tetrahydrofuran isreacted at 140° C. in a single mode microwave (Emrys Optimizer) for 30min. After pre-purification by preparative HPLC (RP18 column; mobilephase: acetonitrile/water gradient), the crude product is taken up inacetonitrile/water/methanol and chromatographed by preparative HPLC(Method 11). The formate salt obtained from the HPLC separation islyophilized and adjusted to pH=7-8 using 1 N aqueous sodium hydroxidesolution. The product is isolated by preparative HPLC (RP18 column;mobile phase: acetonitrile/water gradient). Yield: 23 mg (11% of theory)

LC-MS (Method 4): R_(t)=1.38 min; MS (ESIpos): m/z=315 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.58 (d, 1H), 8.42 (d, 1H), 8.40 (br. s,1H), 7.88 (d, 1H), 7.66 (br. s, 1H), 4.09 (t, 2H), 3.98 (t, 2H),1.86-1.68 (m, 4H).

Example 281-{2-[6-(1,2-Oxazinan-2-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-imidazole-4-carbonitrile

A mixture of 200 mg (1.4 mmol) of the compound from Example 11A, 205 mg(1.7 mmol) of 1,2-oxazinane hydrochloride [Bhat et al., J. Chem. Soc.Perkin Trans. 2 2000, 7, 1435-1446] and 289 μl (215 mg, 1.7 mmol) ofN-ethyl-N-(propan-2-yl)propane-2-amine in 3 ml of water is stirred at100° C. for 1.5 h. Following the addition of 59 μl (49 mg, 0.4 mmol) oftrifluoroacetic acid and 324 mg (1.4 mmol) of the compound from Example6A, the reaction mixture is stirred at 100° C. for 16 h. Theprecipitated solid is filtered off and washed first with water and thenwith diethyl ether. The product is dried under reduced pressure. Yield:199 mg (39% of theory)

LC-MS (Method 5): R_(t)=0.87 min; MS (ESIpos): m/z=339 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=8.57 (d, 1H), 8.45 (d, 1H), 8.40 (br. s,1H), 8.23 (d, 1H), 7.70 (br. s, 1H), 4.07 (t, 2H), 3.96 (t, 2H),1.85-1.65 (m, 4H).

Example 291-{2-[6-(1,2-Oxazinan-2-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-1,2,3-triazole-4-carbonitrile

A mixture of 200 mg (1.4 mmol) of the compound from Example 11A, 205 mg(1.7 mmol) of 1,2-oxazinane hydrochloride [Bhat et al., J. Chem. Soc.Perkin Trans. 2 2000, 7, 1435-1446] and 289 μl (215 mg, 1.7 mmol) ofN-ethyl-N-(propan-2-yl)propane-2-amine in 3 ml of water is stirred at100° C. for 1.5 h. Following the addition of 59 μl (49 mg, 0.4 mmol) oftrifluoroacetic acid and 325 mg (1.4 mmol) of the compound from Example7A, the reaction mixture is stirred at 100° C. for 16 h. Theprecipitated solid is filtered off and washed first with water and thenwith diethyl ether. The product is dried under reduced pressure. Yield:122 mg (24% of theory)

LC-MS (Method 4): R_(t)=1.66 min; MS (ESIpos): m/z=340 [M+H]⁺;

¹H-NMR (400 MHz, DMSO-d₆): δ=9.28 (s, 1H), 8.58 (s, 1H), 8.36 (s, 1H),7.60 (br. s, 1H), 4.12 (t, 2H), 4.03 (t, 2H), 1.88-1.70 (m, 4H).

B. Evaluation of the Pharmacological Activity

The pharmacological properties of the compounds according to theinvention can be demonstrated in the following assays:

Abbreviations: DMEM Dulbecco's modified Eagle medium FCS fetal calfserum TMB 3,3′,5,5′-tetramethylbenzidine Tristris(hydroxymethyl)aminomethane1. In Vitro Tests for Determination of the Activity and Selectivity ofHIF Prolyl 4-hydroxylase Inhibitors1.a) Inhibition of the Activity of HIF Prolyl Hydroxylase:

Hydroxylated HIF bonds specifically to the von Hippel-Lindauprotein-elongin B-elongin C complex (VBC complex). This interactionoccurs only if HIF is hydroxylated on a conserved prolyl radical. It isthe basis for the biochemical determination of HIF prolyl hydroxylaseactivity. The test is carried out as described [Oehme F., Jonghaus W.,Narouz-Ott L., Huetter J., Flamme I., Anal. Biochem. 330 (1), 74-80(2004)]:

A clear 96-well microtiter plate coated with NeutrAvidin HBC (Pierce) isincubated with blocker casein for 30 minutes. The plate is then washedthree times with 200 μl each time of wash buffer (50 mM Tris, pH 7.5,100 mM NaCl, 10% (v/v) blocker casein, 0.05% (v/v) Tween 20) per well.The peptide biotin-DLDLEMLAPYIPMDDDFQL (Eurogentec, 4102 Seraing,Belgium) is added in a concentration of 400 nM in 100 μl wash buffer.This peptide serves as a substrate for the prolyl hydroxylation and isbonded to the microtiter plate. After incubation for 60 minutes, theplate is washed three times with wash buffer, incubated with 1 mM biotinin blocker casein for 30 minutes and then washed again three times withwash buffer.

To carry out the prolyl hydroxylase reaction, the peptide substratebonded to the plate is incubated with a cell lysate containing prolylhydroxylase for 1 to 60 minutes. The reaction takes place in 100 μlreaction buffer (20 mM Tris, pH 7.5, 5 mM KCl, 1.5 mM MgCl₂, 1 μM-1 mM2-oxoglutarate, 10 μM FeSO₄, 2 mM ascorbate) at room temperature. Thereaction mixture moreover contains various concentrations of the prolylhydroxylase inhibitor to be tested. The test substance is preferably,but not exclusively, employed at concentrations of between 1 nM and 100μM. The reaction is stopped by washing the plate three times with washbuffer.

For quantitative determination of the prolyl hydroxylation, a fusionprotein which contains both thioredoxin from E. coli and the VBC complexin 80 μl bonding buffer (50 mM Tris, pH 7.5, 120 mM NaCl) is added.After 15 minutes, 10 μl of a solution of polyclonal anti-thioredoxinantibodies from rabbit in bonding buffer are added. After a further 30minutes, 10 μl of a solution of anti-rabbit immunoglobulin coupled tohorseradish peroxidase in bonding buffer are added. After incubation atroom temperature for 30 minutes, the plate is washed three times withwash buffer in order to remove non-bonded VBC complex and antibodies. Todetermine the amount of bonded VBC complex, the plate is incubated withTMB for 15 minutes. The colour reaction is ended by addition of 100 μl 1M sulphuric acid. The amount of bonded VBC complex is determined bymeasurement of the optical density at 450 nm. It is proportional to theamount of hydroxylated proline in the peptide substrate.

Alternatively, a VBC complex coupled to europium (Perkin Elmer) can beused for detection of the prolyl hydroxylation. In this case, the amountof bonded VBC complex is determined by the fluorescence with respect totime. The use of VBC complex labelled with [³⁵S]-methionine is moreoverpossible. For this, the radioactively labelled VBC complex can beprepared by in vitro transcription-translation in reticulocyte lysate.

The embodiment examples inhibit the activity of HIF prolyl hydroxylasein this test with an IC₅₀ value of ≦30 μM. Representative IC₅₀ valuesfor the embodiment examples are reproduced in the following Table 1:

TABLE 1 Example No. IC₅₀ [nM] 2 880 5 540 9 760 17 130 20 90 21 70 25180 26 380 29 1701.b) Cellular, Functional In Vitro Test:

The activity of the compounds according to the invention is quantifiedwith the aid of a recombinant cell line. The cell is originally derivedfrom a human lung carcinoma cell line (A549, ATCC: American Type CultureCollection, Manassas, Va. 20108, USA). The test cell line is transfectedin a stable manner with a vector which contains the reporter gene ofPhotinus pyralis luciferase (called luciferase in the following) underthe control of an artificial minimal promoter. The minimal promotercomprises two hypoxia-responsible elements upstream of a TATA box [OehmeF., Ellinghaus P., Kolkhof P., Smith T. J., Ramakrishnan S., Hütter J.,Schramm M., Flamme I., Biochem. Biophys. Res. Commun. 296 (2), 343-9(2002)]. Under the effect of hypoxia (e.g. culturing in the presence of1% oxygen for 24 hours) or under the action of non-selective dioxygenaseinhibitors (e.g. desferroxamine in a concentration of 100 μM, cobaltchloride in a concentration of 100 μM or N-oxalylglycine diethyl esterin a concentration of 1 mM), the test cell line produces luciferase,which can be detected and quantified with the aid of suitablebioluminescence reagents (e.g. Steady-Glo® Luciferase Assay System,Promega Corporation, Madison, Wis. 53711, USA) and a suitableluminometer.

Test procedure: On the day before the test, the cells are plated out inan exactly calculated amount of culture medium (DMEM, 10% FCS, 2 mMglutamine) in 384- or 1,536-well microtiter plates and kept in a cellincubator (96% atmospheric humidity, 5% v/v CO₂, 37° C.). On the testday, the test substances are added to the culture medium in graduatedconcentrations. No test substance is added to the cells in batchesserving as negative controls. As a positive control for determination ofthe sensitivity of the cell to inhibitors, e.g. desferroxamine is addedin a final concentration of 100 μM. Six to 24 hours after transfer ofthe test substances into the wells of the microtiter plates, theresulting light signal is measured in the luminometer. A dose/effectrelationship is plotted with the aid of the measurement values, whichserves as a basis for determining the half-maximum active concentration(called the EC₅₀ value).

1.c) Cellular, Functional In Vitro Test of Modification of the GeneExpression:

To investigate the modification of the expression of specific mRNAs inhuman cell lines after treatment with test substances, the followingcell lines are cultured on 6- or 24-well plates: human hepatoma cells(HUH, JCRB Cell Bank, Japan), human embryonal kidney fibroblasts(HEK/293, ATCC, Manassas, Va. 20108, USA), human cervical carcinomacells (HeLa, ATCC, Manassas, Va. 20108, USA), human umbilical veinendothelial cells (HUVEC, Cambrex, East Rutherford, N.J. 07073, USA). 24hours after addition of the test substances, the cells are washed withphosphate-buffered saline and the total RNA is obtained from them usinga suitable method (e.g. Trizol® reagent, Invitrogen GmbH, 76131Karlsruhe, Germany).

For a typical analysis experiment, 1 μg each of the total RNA obtainedin this way is digested with DNase I and translated into a complementaryDNA (cDNA) using a suitable reverse transcriptase reaction (ImProm-IIReverse Transcription System, Promega Corporation, Madison, Wis. 53711,USA). 2.5% of the cDNA batch obtained in this way is used in each casefor the polymerase chain reaction. The expression level of the mRNA ofthe genes to be investigated is investigated by means of the real timequantitative polymerase chain reaction [TaqMan-PCR; Heid C. A., StevensJ., Livak K. J., Williams P. M., Genome Res. 6 (10), 986-94 (1996)]using an ABI Prism 7700 sequence detection instrument (AppliedBiosystems, Inc.). The primer-probe combinations used here are generatedby means of Primer Express 1.5 Software (Applied Biosystems, Inc.).Specifically, the mRNAs of erythropoietin, carboanhydrase IX, lactatedehydrogenase A and vascular endothelial cell growth factor areinvestigated.

Substances according to the present invention lead to a significantdose-dependent increase in the mRNA of hypoxia-induced genes in cells ofhuman origin.

2. In Vivo Tests for Detection of the Action in the CardiovascularSystem

2.a) In Vivo Test of Modification of Gene Expression:

The test compounds dissolved in suitable solvents are administered tomice or rats either orally by stomach tube administration,intraperitoneally or intravenously. Typical dosages are 0.1, 0.5, 1, 5,10, 20, 50, 100 and 300 mg substance per kg of body weight andadministration. Control animals receive only solvent. 4, 8 or 24 hoursafter administration of the test substance the animals are sacrificedwith an overdose of isoflurane and a subsequent fracture of the neck andthe organs to be investigated are removed. Parts of the organs areshock-frozen in liquid nitrogen. Total RNA is obtained from the organparts as described under B.1.a) and this is translated into a cDNA. Theexpression level of the mRNA of the genes to be investigated isinvestigated by means of the real time quantitative polymerase chainreaction [TaqMan-PCR; Heid C. A., Stevens J., Livak K. J., Williams P.M., Genome Res. 6 (10), 986-94 (1996)] using an ABI Prism 7700 sequencedetection instrument (Applied Biosystems, Inc.).

Substances according to the present invention lead to a significantdose-dependent increase in the mRNA of erythropoietin in the kidneyafter oral or parenteral administration compared with the placebocontrol.

2.b) Determination of the Erythropoietin Level in Serum:

The test substance in a suitable solvent is administered to mice or ratseither intraperitoneally or orally once or twice daily. Typical dosagesare 0.1, 0.5, 1, 5, 10, 20, 50, 100 and 300 mg substance per kg of bodyweight and administration. Placebo control animals receive only solvent.Before the administration and four hours after the last administrationof substance, 50 μl of blood are taken from the animals from theretroorbital venous plexus or the tail vein under short narcosis. Theblood is rendered uncoagulable by addition of lithium heparin. The bloodplasma is obtained by centrifugation. The content of erythropoietin inthe blood plasma is determined with the aid of an erythropoietin-ELISA(Quantikine® mouse Epo Immunoassay, R&D Systems, Inc., Minneapolis, USA)in accordance with the manufacturer's instructions. The measurementvalues are converted into pg/ml with the aid of a reference measurementrecorded for mouse erythropoietin.

Substances according to the present invention lead to a significantdose-dependent increase in the plasma erythropoietin after oral andparental administration compared with the starting value and the placebocontrol.

2.c) Determination of the Cell Composition of Peripheral Blood:

The test substance in a suitable solvent is administered to mice or ratseither intraperitoneally or orally once or twice daily for several days.Typical dosages are e.g. 0.1, 0.5, 1, 5, 10, 20, 50, 100 and 300 mgsubstance per kg of body weight and administration. Control animalsreceive only solvent. At the end of the study, blood is taken from theanimals from the venous plexus of the corner of the eye or the tail veinunder short narcosis and is rendered uncoagulable by addition of sodiumcitrate. The concentrations of erythrocytes, leukocytes and thrombocytesare determined in the blood samples in a suitable electronic measuringapparatus. The concentration of the reticulocytes is determined bymicroscope screening of in each case 1000 erythrocytes with the aid ofblood smears stained with a stain solution suitable for this purpose(KABE Labortechnik, Nümbrecht). For determination of the haematocrit,blood is taken from the retroorbital venous plexus by means of ahaematocrit capillary and the haematocrit value is read off manuallyafter centrifugation of the capillary in a centrifuge suitable for thispurpose.

Substances according to the present invention lead to a significantdose-dependent increase in the haematocrit, the erythrocyte count andthe reticulocytes after oral and parenteral administration compared withthe starting value and the placebo control.

3. Determination of the Solubility

Preparation of the Starting Solution (Initial Solution):

At least 1.5 mg of the test substance are weighed out accurately into aWide Mouth 10 mm Screw V-Vial (from Glastechnik Grädfenroda GmbH, Art.No. 8004-WM-H/V15μ) with fitting screw cap and septum, DMSO is added togive a concentration of 50 mg/ml and the mixture is vortexed for 30minutes.

Preparation of the Calibration Solutions:

The required pipetting steps are carried out in a 1.2 ml Deep Well Plate(DWP) with 96 wells using a liquid handling robot. The solvent used is amixture of acetonitrile/water 8:2.

Preparation of the starting solution for calibration solutions (stocksolution): 833 μl of the solvent mixture are added to 10 μl of theinitial solution (concentration=600 μg/ml), and the mixture ishomogenized. For each test substance, 1:100 dilutions are prepared inseparate DWPs, and the dilutions for their part are homogenized.

Calibration solution 5 (600 ng/ml): 270 μl of solvent mixture are addedto 30 μl of the stock solution, and the mixture is homogenized.

Calibration solution 4 (60 ng/ml): 270 μl of solvent mixture are addedto 30 μl of calibration solution 5, and the mixture is homogenized.

Calibration solution 3 (12 ng/ml): 400 μl of solvent mixture are addedto 100 μl of calibration solution 4, and the mixture is homogenized.

Calibration solution 2 (1.2 ng/ml): 270 μl of solvent mixture are addedto 30 μl of calibration solution 3, and the mixture is homogenized.

Calibration solution 1 (0.6 ng/ml): 150 μl of solvent mixture are addedto 150 μl of calibration solution 2, and the mixture is homogenized.

Preparation of the Sample Solutions:

The required pipetting steps are carried out in a 1.2 ml DWP with 96wells using a liquid handling robot. 1000 μl of PBS buffer pH 6.5 areadded to 10.1 μl of the stock solution. (PBS buffer pH 6.5: 61.86 g ofsodium chloride, 39.54 g of sodium dihydrogen phosphate and 83.35 g of 1N aqueous sodium hydroxide solution are weighed out into a 1 litremeasuring flask, the flask is filled with water and the mixture isstirred for about 1 hour. From this solution, 500 ml are added to a5-litre measuring flask, and the flask is filled with water. Using 1 Naqueous sodium hydroxide solution, the pH is adjusted to 6.5.)

Practice:

The required pipetting steps are carried out in a 1.2 ml DWP with 96wells using a liquid handling robot. Using a temperature-adjustableshaker, the sample solutions prepared in this manner are shaken at 20°C. and 1400 rpm for 24 hours. From these solutions, in each case 180 μlare removed and transferred into Beckman polyallomer centrifuge tubes.These solutions are centrifuged at about 223 000×g for 1 hour. From eachsample solution, 100 μl of the supernatant are removed and diluted 1:10and 1:1000 with PBS buffer 6.5.

Analysis:

The samples are analyzed by HPLC/MS-MS. Quantification is carried outusing a five-point calibration curve of the test compound. Thesolubility is expressed in mg/l. Analysis sequence: 1) blank (solventmixture); 2) calibration solution 0.6 ng/ml; 3) calibration solution 1.2ng/ml; 4) calibration solution 12 ng/ml; 5) calibration solution 60ng/ml; 6) calibration solution 600 ng/ml; 7) blank (solvent mixture); 8)sample solution 1:1000; 7) sample solution 1:10.

HPLC/MS-MS Method

HPLC: Agilent 1100, quat. pump (G1311A), autosampler CTC HTS PAL,degasser (G1322A) and column thermostat (G1316A); column: Oasis HLB 20mm×2.1 mm, 25 p; temperature: 40° C.; mobile phase A: water+0.5 ml offormic acid/l; mobile phase B: acetonitrile+0.5 ml of formic acid/l;flow rate: 2.5 ml/min; stop time 1.5 mm; gradient: 0 mm 95% A, 5% B;ramp: 0-0.5 mm 5% A, 95% B; 0.5-0.84 min 5% A, 95% B; ramp: 0.84-0.85min 95% A, 5% B; 0.85-1.5 min 95% A, 5% B.

MS/MS: WATERS Quattro Micro Tandem MS/MS; Z-Spray API interface; HPLC-MSinitial splitter 1:20; measurement in the ESI mode.

C. Embodiment Examples for Pharmaceutical Compositions

The compounds according to the invention can be converted intopharmaceutical formulations as follows:

Tablet:

Composition:

100 mg of the compound according to the invention, 50 mg lactose(monohydrate), 50 mg maize starch (native), 10 mg polyvinylpyrrolidone(PVP 25) (BASF, Ludwigshafen, Germany) and 2 mg magnesium stearate.

Tablet weight 212 mg. Diameter 8 mm, radius of curvature 12 mm.

Preparation:

The mixture of compound according to the invention, lactose and starchis granulated with a 5% strength solution (w/w) of the PVP in water.After drying, the granules are mixed with the magnesium stearate for 5minutes. This mixture is pressed with a conventional tablet press (fortablet format see above). A pressing force of 15 kN is used as therecommended value for the pressing.

Suspension for Oral Administration:

Composition:

1000 mg of the compound according to the invention, 1000 mg ethanol(96%), 400 mg Rhodigel® (xanthan gum from FMC, Pennsylvania, USA) and 99g water.

10 ml of oral suspension correspond to an individual dose of 100 mg ofthe compound according to the invention.

Preparation:

The Rhodigel is suspended in ethanol and the compound according to theinvention is added to the suspension. The water is added with stirring.The mixture is stirred for approx. 6 h until swelling of the Rhodigelhas ended.

Solution for Oral Administration:

Composition:

500 mg of the compound according to the invention, 2.5 g polysorbate and97 g polyethylene glycol 400.20 g of oral solution correspond to anindividual dose of 100 mg of the compound according to the invention.

Preparation:

The compound according to the invention is suspended in a mixture ofpolyethylene glycol and polysorbate, while stirring. The stirringoperation is continued until dissolution of the compound according tothe invention is complete.

i.v. Solution:

The compound according to the invention is dissolved in a concentrationbelow the saturation solubility in a physiologically acceptable solvent(e.g. isotonic saline solution, glucose solution 5% and/or PEG 400solution 30%). The solution is subjected to sterile filtration and istransferred into sterile and pyrogen-free injection containers.

1. A compound of the formula

in which X represents N or CH; R¹ represents hydrogen or cyano; and R²represents a saturated 4- to 7-membered heterocyclyl radical which isattached via a nitrogen atom, where the heterocyclyl radical may besubstituted by a substituent, where the substituent is selected from thegroup consisting of hydroxyl, hydroxycarbonyl, C₁-C₃-alkyl,C₁-C₃-alkylamino and C₃-C₆-cycloalkyl, or where the heterocyclyl radicalmay be substituted by 1 to 4 fluorine substituents, or a salt thereof.2. A compound according to claim 1, wherein X represents N or CH; R¹represents hydrogen or cyano; and R² represents a saturated 4- to7-membered heterocyclyl radical which is attached via a nitrogen atom,where the heterocyclyl radical is substituted by 1 to 4 fluorinesubstituents, or R² represents piperazin-1-yl, where piperazin-1-yl issubstituted by one substituent, where the substituent is selected fromthe group consisting of C₃-C₆-cycloalkyl, or R² representsazetidin-1-yl, where azetidin-1-yl is substituted by one substituent,where the substituent is selected from the group consisting ofhydroxycarbonyl, C₁-C₃-alkyl, C₁-C₃-alkylamino and C₃-C₆-cycloalkyl, orR² represents 1,2-oxazinan-2-yl or 1,4-oxazepan-4-yl, or a salt thereof.3. A compound according to claim 1, wherein X represents N or CH; R¹represents hydrogen or cyano; and R² represents azetidin-1-yl,pyrrolin-1-yl or piperidin-1-yl, where azetidin-1-yl, pyrrolin-1-yl andpiperidin-1-yl are substituted by 1 to 4 fluorine substituents, or R²represents piperazin-1-yl, where piperazin-1-yl is substituted in the4-position by one substituent, where the substituent is selected fromthe group consisting of C₃-C₆-cycloalkyl, or R² representsazetidin-1-yl, where azetidin-1-yl is substituted in the 3-position byone substituent, where the substituent is selected from the groupconsisting of hydroxycarbonyl, methyl and dimethylamino, or R²represents 1,2-oxazinan-2-yl or 1,4-oxazepan-4-yl, or a salt thereof. 4.A method for preparing a compound of the formula (I) or a salt thereofaccording to claim 1, wherein [A] a compound of the formula

in which R¹ has the meaning given in claim 1, and Z¹ represents methylor ethyl, is reacted in an inert solvent, if appropriate in the presenceof an acid, with a compound of the formula

in which R² has the meaning given in claim 1, to give a compound of theformula

in which Z¹, R¹ and R² have the meaning given in claim 1, which, alreadyunder these reaction conditions or in a subsequent reaction step underthe influence of a base, cyclizes to give the compound of the formula(I), and the compound of the formula (I) is, if appropriate with theappropriate (i) solvent and/or (ii) base or acid, converted into one ofits salts, or [B] a compound of the formula

in which Z¹ and R¹ have the meaning given in claim 1, is condensed witha compound of the formula

in which Z² represents methyl or ethyl, to give a compound of theformula

in which Z¹ and R¹ have the meaning given in claim 1, and then reactedin the presence of an acid with a compound of the formula (III) to givea compound of the formula (IV), which, already under these reactionconditions or in a subsequent reaction step under the influence of abase, cyclizes to give the compound of the formula (I), and the compoundof the formula (I) is, if appropriate with the appropriate (i) solventand/or (ii) base or acid, converted into one of its salts, or [C] thecompound of the formula

is, in water as the solvent in a one-pot process, reacted initially witha compound of the formulaR¹—H  (IX), in which R² has the meaning given in claim 1, and then witha compound of the formula (VII) to give a compound of the formula (I),and the compound of the formula (I) is, if appropriate with theappropriate (i) solvent and/or (ii) base or acid, converted into one ofits salts.
 5. A pharmaceutical composition comprising a compoundaccording to claim 1 in combination with an inert, non-toxic,pharmaceutically acceptable excipient.
 6. A method for treating anemiain a subject comprising administering a therapeutically effective amountof a compound according to claim
 1. 7. A compound according to claim 1,characterized in that X represents N or CH, R¹ represents hydrogen orcyano, and R² represents piperazin-1-yl, wherein piperazin-1-yl issubstituted by a substituent selected from the group consisting ofC₃-C₆-cycloalkyl, or a salt thereof.
 8. A compound according to claim 7,characterized in that X represents N or CH, R¹ represents hydrogen orcyano, and R² represents piperazin-1-yl, wherein piperazin-1-yl issubstituted in the 4-position by a substituent selected from the groupconsisting of C₃-C₆-cycloalkyl, or a salt thereof.
 9. A compoundaccording to claim 1, characterized in that X represents N or CH, R¹represents hydrogen or cyano, and R² represents azetidin-1-yl, whereinazetidin-1-yl is substituted by a substituent selected from the groupconsisting of hydroxycarbonyl, C₁-C₃-alkyl, C₁-C₃-alkylamino andC₃-C₆-cycloalkyl, or a salt thereof.
 10. A compound according to claim9, characterized in that X represents N or CH, R¹ represents hydrogen orcyano, and R² represents azetidin-1-yl, wherein azetidin-1-yl issubstituted by a substituent selected from the group consisting ofhydroxycarbonyl, methyl and dimethylamino, or a salt thereof.
 11. Acompound according to claim 10, characterized in that X represents N orCH, R¹ represents hydrogen or cyano, and R² represents azetidin-1-yl,wherein azetidin-1-yl is substituted in the 3-position by a substituentselected from the group consisting of hydroxycarbonyl, methyl anddimethylamino, or a salt thereof.
 12. A compound according to claim 1,characterized in that X represents N or CH, R¹ represents hydrogen orcyano, and R² represents 1,2-oxazinan-2-yl or 1,4-oxazepan-4-yl, or asalt thereof.
 13. A compound according to claim 1, wherein R² represents4-cyclobutyl-piperazin-1-yl.
 14. A compound according to claim 1,wherein the compound is1-{2-[6-(4-Cyclobutylpiperazin-1-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-imidazole-4-carbonitrile,as represented by the formula

or a salt thereof.
 15. A compound according to claim 1, where thecompound is1-{2-[6-(4-Cyclobutylpiperazin-1-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-imidazole-4-carbonitrile,as represented by the formula


16. A compound according to claim 1, where the compound is1-{6-[5-Oxo-4-(1H-1,2,3-triazol-1-yl)-2,5-dihydro-1H-pyrazol-1-yl]pyrimidin-4-yl}azetidine-3-carboxylicacid, as represented by the formula

or a salt thereof.
 17. A compound according to claim 1, where thecompound is1-{6-[5-Oxo-4-(1H-1,2,3-triazol-1-yl)-2,5-dihydro-1H-pyrazol-1-yl]pyrimidin-4-yl}azetidine-3-carboxylicacid, as represented by the formula


18. A compound according to claim 1, where the compound is2-[6-(1,2-Oxazinan-2-yl)pyrimidin-4-yl]-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one,as represented by the formula

or a salt thereof.
 19. A compound according to claim 1, where thecompound is 2-[6-(1,2-Oxazinan-2-yl)pyrimidin-4-yl]-4-(1H-1,2,3-triazol-1-yl)-1,2-dihydro-3H-pyrazol-3-one,as represented by the formula


20. A compound according to claim 1, where the compound is1-{2-[6-(1,2-Oxazinan-2-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-1,2,3-triazole-4-carbonitrile,as represented by the formula:

or a salt thereof.
 21. A compound according to claim 1, wherein thecompound is1-{2-[6-(1,2-Oxazinan-2-yl)pyrimidin-4-yl]-3-oxo-2,3-dihydro-1H-pyrazol-4-yl}-1H-1,2,3-triazole-4-carbonitrile,as represented by the formula: